Patent Abstract:
The present invention provides methods and materials useful for monitoring and regulating the glycosylation of glycoproteins that are recombinantly produced from cells. In particular, methods are provided for monitoring and regulating levels of cellular indicators which affect the level of fucosylation produced by cells.

Full Description:
CLAIM OF PRIORITY 
       [0001]    This application claims priority under 35 USC §119(e) to U.S. Patent Application Ser. No. 61/184,493, filed on Jun. 5, 2009, the entire contents of which is hereby incorporated by reference. 
     
    
     FIELD OF THE INVENTION 
       [0002]    The invention relates to glycoproteins and glycoprotein preparations having reduced core fucosylation and methods related thereto, e.g., methods of making and using the glycoproteins and glycoprotein preparations. 
       BACKGROUND OF INVENTION 
       [0003]    A typical glycoprotein consists not only of an amino acid backbone but also includes one or more glycan moieties. The glycan moieties attached to the amino acid backbone of a glycoprotein can vary structurally in many ways including, sequence, branching, sugar content, and heterogeneity. Glycosylation adds not only to the structural complexity of the molecules, but also affects or conditions many of a glycoprotein&#39;s biological and clinical attributes. 
       SUMMARY OF INVENTION 
       [0004]    As is disclosed herein, the relationship between GDP-fucose levels in a cell and the level of fucosylation of proteins produced by a cell is not linear. A relatively modest reduction in GDP-fucose levels in the cell can result in a much lower level of fucosylation on proteins produced by the cell. Thus, when levels of GDP-fucose taught herein are used, the reduction of fucose on proteins produced by the cells can be maximized with minimal reduction in GDP-fucose levels and minimal disruption of other aspects of metabolism. E.g., one or more manipulations described herein can be used to achieve a minimal reduction of GDP-fucose levels but still provide a relatively great reduction in fucosylation. Thus, methods described herein allow optimization of the levels of GDP-fucose reduction with reduction in the fucosylation of proteins made by the cell. 
         [0005]    The inventors have shown that the relationship between the level of GDP-fucose in a cell and the level of fucosylation on proteins made by the cell is non-linear. In embodiments the curve which describes the relationship between level of GDP-fucose in a cell and level of fucosylation of proteins made by the cell includes three phases. In embodiments the three phase are as follows: a first phase, beginning at relatively high concentrations of GDP-fucose, and continuing through declining levels of GDP-fucose, wherein the level of fucosylation on proteins made by the cell is, compared to the other two phases, relatively constant; a second phase, beginning at levels of GDP-fucose that are lower than the levels seen in the first phase, wherein the level of fucosylation on proteins made by the cell, compared to the other two phases, drops rapidly in response to a decrease in GDP-fucose level; and a third phase, beginning at levels of GDP-fucose that are lower than levels in the second phase, and continuing through declining levels of GDP-fucose, wherein the level of fucosylation on proteins made by the cell is, compared to the other two phases, relatively constant. 
         [0006]    In embodiments the curve which describes the relationship between level of GDP-fucose in a cell and level of fucosylation of proteins made by the cell has three phases: a phase having a high relatively constant (relatively independent of the amount of GDP-fucose) level of fucosylation (points to the left of point A in  FIG. 1 ), a phase of rapid decrease in fucosylation (points between A and B in  FIG. 1 , wherein the level of fucosylation is relatively sensitive to the amount of GDP-fucose), and phase having a lower, relatively constant, level of fucosylation (relatively independent of the amount of GDP-fucose) (points to the right of point B in  FIG. 1 ). ( FIG. 1  and the contents therein are typical. Of course analogous plots may also be used. In embodiments the curve plotting the relationship between level of GDP-fucose in a cell and level of fucosylation of proteins made by the cell may look different from that in  FIG. 1 , but it will still have the three phases described.) 
         [0007]    The appreciation of this relationship can be used to guide selection of the level of GDP-fucose, e.g., to allow minimization of the level of fucosylation with minimal reduction in the level of GDP-fucose in the cell. The balance between low fucose and undesirable contributions of low GDP-fucose levels can be optimized. This can allow minimizing the negative effects of very low concentrations of GDP-fucose. 
         [0008]    For example, in some embodiments a decrease in GDP-mannose concentrations can be an undesirable side effect of very low GDP-fucose levels. In some instances a loss of GDP-fucose can lead to higher levels of conversion of GDP-mannose to GDP-fucose, leading to an undesirable decrease in intracellular levels of GDP-mannose. A decrease in GDP-mannose can result in a decrease in high mannose structures on proteins produced by the cell. High mannose structures mediate effector function, and particularly ADCC activity, of an antibody. Thus, if ADCC activity is a desirable property, a decrease in high mannose structures can be undesirable. Alternatively, if less ADCC activity is desired decreased GDP-mannose can be desirable. 
         [0000]    Optimal levels can be determined by monitoring the levels of GDP-mannose in the cell; as needed the levels of GDP-fucose can be elevated if the levels of GDP-mannose begin to drop. In particular embodiments, GDP-fucose is increased, e.g., added, if GDP-mannose levels are less than about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 25%, 20%, 15% or 10% of a reference GDP-mannose level, e.g., the level seen in an otherwise similar cell that does not have a reduction in GDP-mannose. 
         [0009]    In other embodiments an increase in GDP-mannose concentrations is can be an undesirable side effect of very low GDP-fucose levels. In some instances a loss of GDP-fucose may lead to decreased conversion of GDP-mannose to GDP-fucose, leading to an undesirable increase in the levels of GDP-mannose (in some embodiments this might be observed when a cell is largely or completely deficient in the enzymes involved in the conversion of GDP-mannose to GDP-fucose). Optimal levels can be determined by monitoring the levels of GDP-mannose in the cell; as needed the levels of GDP-fucose or the level of the converting enzyme responsible for the GDP-fucose can be elevated if the levels of GDP-mannose begin to rise. In particular embodiments, GDP-fucose or the level of the converting enzyme is increased if GDP-mannose levels are more than about 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference GDP-mannose level, e.g. the level seen in an otherwise similar cell that does not have reduction ion the GDP-mannose. 
         [0010]    The invention features glycoproteins, e.g., antibodies, and preparations thereof having reduced fucosylation, e.g., reduced core fucosylation. Exemplary proteins include a peptide which comprises a human IgG constant region, e.g., one made in cultured cells, e.g., CHO cells, and having a glycan component attached in the CH2 region, e.g., at residue Asn 297. Preparations, e.g., pharmaceutically acceptable preparations, of these, and other proteins having reduced levels of fucosylation, e.g., core fucosylation, are provided. The presence of core fucosylation on an antibody significantly attenuates its ADCC activity. Reduction of core fucosylation increases ADCC activity. 
         [0011]    The invention provides methods in which cells having a manipulation (defined below) can be used to provide proteins having reduced fucosylation. E.g, one or both of a genetically engineered alteration and culture conditions can be used to provide an optimized level of GDP-fucose and an optimized level of fucosylation on proteins made by a cell. 
         [0012]    Accordingly, in one aspect, the invention features, a method of reducing fucosylation of a glycoprotein (or a preparation of glycoproteins). The method comprises: 
         [0013]    providing a cell having or subject to a manipulation that results in a level of GDP-fucose in said cell that is below a first preselected level and, in embodiments, above a second preselected level and optionally memorializing one or both levels; 
         [0014]    culturing said cell, e.g., to provide a batch of cultured cells; 
         [0015]    optionally, measuring the level of GDP-fucose in said cell or batch of cultured cells; 
         [0016]    optionally, separating the glycoprotein from at least one component with which said cell or batch of cultured cells was cultured; and 
         [0017]    optionally, evaluating the glycoprotein (or a glycoprotein on the surface of the cell) for a parameter related to fucosylation; 
         [0018]    thereby providing a glycoprotein with reduced fucosylation, e.g., wherein the level of fucosylation is reduced by a predetermined level in comparison with a reference. 
         [0019]    In an embodiment the manipulation is or was selected on the basis of providing a level of GDP fucose below a first preselected level and optionally above a second preselected level. 
         [0020]    In one embodiment, the method further comprises evaluating a glycan on the surface of said cell or batch of cultured cells in order to determine if the glycoprotein produced by said cell or batch of cultured cells has reduced fucosylation. In another embodiment, said evaluation comprises evaluating a glycan on the surface of said cell or batch of cultured cells, to determine a property of said glycan, comparing the property to a reference, to thereby determine if said glycan structure is present on the product. 
         [0021]    In one embodiment, said first preselected level of GDP-fucose is selected from a level that is: 
         [0022]    i.a) approximately equal to or less than 80%, 70% or 60% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0023]    ii.a) approximately equal to, or less than, the point of maximum curvature above the inflection point (e.g., the inflection point in the second phase) on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0024]    ii.1.a) approximately equal to, or less than, the lowest level that results in a normal (e.g., that seen in an un-manipuated cell) level of fucosylation; 
         [0025]    iii.a) approximately equal to or less than the point of maximum curvature below the inflection point on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0026]    iii.1.a) approximately equal to, or less than, the highest level that results in no further reduction in fucosylation; 
         [0027]    iv.a) approximately equal to or less than point A on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; 
         [0028]    v.a) approximately equal to or less than that corresponding to an amount between points A and B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; or 
         [0029]    vi.a) approximately equal to or less than point B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0030]    In one embodiment, said second preselected level of GDP-fucose is selected from a level: 
         [0031]    i.b) approximately equal to, or greater than, 10%, 15%, 20%, 25%, 30%, 35% or 40% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0032]    ii.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in decrease of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, greater than, 10%, 20%, 30%, 40% or 50% than a reference levee, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0033]    iii.b) an amount that provides an unacceptable level of fucose deprivation, e.g. an amount that results in a level of high mannose structures that are less than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of a reference level 
         [0034]    iv.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of GDP-mannose, e.g. an increase in GDP-mannose that is equal to or greater than 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level, e.g. the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0035]    v.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of high mannose structures that are more than or equal to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level; 
         [0036]    vi.b) approximately equal to or greater than point C on the curve in  FIG. 1 , or greater than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0037]    In an embodiment the first level is i.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0038]    In an embodiment the first level is ii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0039]    In an embodiment the first level is ii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0040]    In an embodiment the first level is iii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0041]    In an embodiment the first level is iii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0042]    In an embodiment the first level is iv.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0043]    In an embodiment the first level is v.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0044]    In an embodiment the first level is vi.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0045]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is i.b. 
         [0046]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is ii.b. 
         [0047]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iii.b. 
         [0048]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iv.b. 
         [0049]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is v.b. 
         [0050]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is vi.b. 
         [0051]    In an embodiment the level of GDP-fucose is between point B and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0052]    In an embodiment the level of GDP-fucose is between point A and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0053]    In one embodiment, the level of GDP-fucose is selected to be outside the range between A and B on the curve in  FIG. 1  (as relatively small changes in GDP-fucose will result in relatively large changes in the amount of fucosylation. In an embodiment the level is also less than B.) In another embodiment, the level of GDP-fucose is reduced by a predetermined level, e.g., in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell having the manipulation. In another embodiment, the level of GDP-fucose is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0054]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the amount of fucosylation in the glycan complement, the amount or fucosylation on a component of the glycan complement, or the amount of fucosylation on a glycan component, e.g., in a preparation of glycoproteins. 
         [0055]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the proportion of a preselected glycan component which bears a fucosyl moiety, e.g., at a selected position on the glycan component, e.g., in a preparation of glycoproteins. 
         [0056]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. In another embodiment, the level of fucosylation is reduced by a predetermined level in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell or batch of cultured cells having the manipulation. In another embodiment, the level of fucosylation is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0057]    In one embodiment, X F  is greater than X G , 
         [0058]    and wherein, 
         [0059]    X F  is the % or proportion of reduction in the level of fucosylation (e.g., as compared to the level of fucosylation in a cell or batch of cultured cells lacking the manipulation); and 
         [0060]    X G  is the % or proportion of reduction in the level of GDP fucose (as compared to the level of GDP fucose in a cell or batch of cultured cells lacking the manipulation). 
         [0061]    In one embodiment, said manipulation is not a genetic lesion or the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety. For example, the manipulation is not a lesion that decreases the expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells is wild-type for one or all of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells does not include an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, absent the manipulation, the level of fucosylation is substantially the same as the level in a wild-type cell. In another embodiment, the manipulated cell carries no mutation that substantially lowers GDP-fucose levels. In another embodiment, the manipulated cell has no siRNA that substantially lowers GDP-fucose levels. 
         [0062]    In one embodiment, the cell has a mutation (e.g., a genetically engineered change) that decreases the level of GDP-fucose. Exemplary mutations include those which alter the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0000]    The mutation can be in the structural gene which encodes GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. Such mutations can decrease the activity of the encoded protein. The decrease can be partial or complete. Such mutations can act, e.g., by altering the catalytic activity of the protein or by altering its half-life. Other exemplary mutations can be in a sequence that control expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. These can be mutations that completely, or partially, reduce the expression of the gene, at the RNA or protein level. Such mutations include deletion or other mutations in endogenous of control sequence. Such mutations also include the introduction of heterologous control sequence, e.g., the introduction of heterologous control regions, e.g., a sequence that will give a desired level of expression. (A heterologous control sequence is a sequence other than a sequence naturally associated with and operably linked to the structural gene.) In embodiments the manipulation comprises a mutation in the structural region or in a control sequence operably linked to the gene. 
         [0063]    In an embodiment a cell having a mutation that that decreases the level of GDP-fucose, e.g., a mutation that decreases the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter is cultured in the presence of a substance, e.g., fucose, that results in a GDP-fucose level and/or a fucosylation level described herein. In an embodiment the cell includes a mutation that, in the absence of fucose in the culture medium, would result in a cell having an unacceptably low level of GDP-fucose. When, however, cultured under the appropriate conditions, e.g., media supplemented, e.g., with fucose, that cell can exhibit a desired level of GDP-fucose, e.g., a level of GDP-fucose described herein. Thus, fucose or another substance is present in the culture medium at a level that results in a level of GDP-fucose recited above. 
         [0064]    In another embodiment, the manipulation is the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety, e.g., an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter, and fucose or another substance is present in the culture medium at a level that results in a level of GDP-fucose recited above. 
         [0065]    In one embodiment, said culturing comprises culturing the cell in a medium that results in said level of GDP-fucose. 
         [0066]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0067]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0068]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0069]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0070]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0071]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0072]    In one embodiment, the manipulation is, or is the product of, a selection for reduced levels of GDP-fucose. In another embodiment, the manipulation is, or is the product of, a selection for reduced fucosylation of a glycoprotein. In another embodiment, the manipulation comprises contact with, or inclusion in or on the cell or batch of cultured cells, of an exogenous inhibitor of an enzyme involved in GDP-fucose biosynthesis, e.g., a specific or non-specific inhibitor. 
         [0073]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. 
         [0074]    In one embodiment, one or more of said cell or said batch of cultured cells, said manipulation, and said glycoprotein, is selected on the basis that it or the combination will provide a glycoprotein having reduced fucosylation. 
         [0075]    In one embodiment, one or more of said cell or said batch of cultured cells, said manipulation (or manipulations), and said glycoprotein, is selected on the basis that it or the combination will provide a level of GDP-fucose described herein, e.g., a level which gives a minimal level of fucosylation (e.g., with reference to a curve analogous to that in  FIG. 1 , the level is to the right of point B) but which is above a preselected level In some embodiments the level is above a level that gives an unwanted decrease in the level of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, or more than, 10%, 20%, 30%, 40% or 50% as compared to a reference level, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation. 
         [0076]    In some embodiments the level is above a level that gives an unwanted increase in the level of GDP-mannose, e.g., an increase in GDP-mannose that is equal to, or more than, about 2×, 3×, 4×, 5×, ×, 7×, 8×, 9×, or 10× of a reference level, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation. 
         [0077]    In one embodiment, the method further comprises providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose. 
         [0000]    In another embodiment, the method further comprises providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below the second level. In another embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In one embodiment, the compound other than GDP-fucose is GDP-mannose. In one embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0078]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In one embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0079]    In an embodiment, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. The inhibitor can be used with a cell which is mutant or wildtype for one or more GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0080]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. Such media can be used, e.g., with a cell having a mutation that eliminates or decreased the activity of one or more of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0081]    While some methods described herein rely at least in part on mutations in a gene that conditions the level of GDP-fucose other methods described herein do not. Thus, cells that are not mutant at key genes involved in maintaining GDP-fucose levels can be used to provide proteins having reduced fucosylation. Levels of GDP-fucose can, e.g., be manipulated by culture conditions. 
         [0082]    Thus, in another aspect, the invention features, a method of reducing fucosylation of a glycoprotein or a preparation of glycoproteins, the method comprising: 
         [0083]    providing a cell that expresses said glycoprotein and that is wild-type for one or more (or all) of GMD, FX, fucokinase, GFPP, GDP-Fucose synthetase, a fucosyltransferase or a GDP-Fucose transporter; 
         [0084]    culturing said cell under conditions that result in a level of GDP-fucose in said cell that is below a first preselected level and, in embodiments, above a second preselected level, and results in a preselected level of fucosylation, which is less than in a reference cell cultured under reference conditions, e.g., to provide a batch of cultured cells; 
         [0085]    optionally, measuring the level of GDP-fucose in said cell or batch of cultured cells; and 
         [0086]    optionally, separating the glycoprotein from at least one component with which said cell or batch of cultured cells was cultured, 
         [0087]    optionally, evaluating the glycoprotein (or a glycoprotein on the surface of the cell or batch of cultured cells) for a parameter related to fucosylation; 
         [0088]    thereby providing a glycoprotein with reduced fucosylation, e.g., wherein the level of fucosylation is reduced by a predetermined level in comparison with a reference. 
         [0089]    In one embodiment, the method further comprises evaluating a glycan on the surface of said cell or batch of cultured cells in order to determine if the glycoprotein produced by said cell or batch of cultured cells has reduced fucosylation. In another embodiment, said evaluation comprises evaluating a glycan on the surface of said cell or batch of cultured cells, to determine a property of said glycan, comparing the property to a reference, to thereby determine if said glycan structure is present on the product. 
         [0090]    In one embodiment, said first preselected level of GDP-fucose is selected from a level that is: 
         [0091]    i.a) approximately equal to or less than 80%, 70% or 60% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0092]    ii.a) approximately equal to, or less than, the point of maximum curvature above the inflection point (e.g., the inflection point in the second phase) on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0093]    ii.1.a) approximately equal to, or less than, the lowest level that results in a normal (e.g., that seen in an un-manipuated cell) level of fucosylation; 
         [0094]    iii.a) approximately equal to or less than the point of maximum curvature below the inflection point on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0095]    iii.1.a) approximately equal to, or less than, the highest level that results in no further reduction in fucosylation; 
         [0096]    iv.a) approximately equal to or less than point A on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; 
         [0097]    v.a) approximately equal to or less than that corresponding to an amount between points A and B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; or 
         [0098]    vi.a) approximately equal to or less than point B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0099]    In one embodiment, said second preselected level of GDP-fucose is selected from a level: 
         [0100]    i.b) approximately equal to, or greater than, 10%, 15%, 20%, 25%, 30%, 35% or 40% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0101]    ii.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in decrease of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, greater than, 10%, 20%, 30%, 40% or 50% than a reference levee, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0102]    iii.b) an amount that provides an unacceptable level of fucose deprivation, e.g. an amount that results in a level of high mannose structures that are less than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of a reference level; 
         [0103]    iv.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of GDP-mannose, e.g. an increase in GDP-mannose that is equal to or greater than 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level, e.g. the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0104]    v.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of high mannose structures that are more than or equal to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level; or 
         [0105]    vi.b) approximately equal to or greater than point C on the curve in  FIG. 1 , or greater than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0106]    In an embodiment the first level is i.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0107]    In an embodiment the first level is ii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0108]    In an embodiment the first level is ii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0109]    In an embodiment the first level is iii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0110]    In an embodiment the first level is iii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0111]    In an embodiment the first level is iv.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0112]    In an embodiment the first level is v.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0113]    In an embodiment the first level is vi.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0114]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is i.b. 
         [0115]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is ii.b. 
         [0116]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iii.b. 
         [0117]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iv.b. 
         [0118]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is v.b. 
         [0119]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is vi.b. 
         [0120]    In an embodiment the level of GDP-fucose is between point B and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0121]    In an embodiment the level of GDP-fucose is between point A and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0122]    In one embodiment, the level of GDP-fucose is selected to be outside the range between A and B on the curve in  FIG. 1  (as relatively small changes in GDP-fucose will result in relatively large changes in the amount of fucosylation. In an embodiment the level is also less than B.). In another embodiment, the level of GDP-fucose is reduced by a predetermined level, e.g., in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, cultured under reference conditions but otherwise the same or essentially the same as the cell cultured under conditions that result in said level of GDP-fucose. In another embodiment, the level of GDP-fucose is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0123]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the amount of fucosylation in the glycan complement, the amount or fucosylation on a component of the glycan complement, or the amount of fucosylation on a glycan component, e.g., in a preparation of glycoproteins. 
         [0124]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the proportion of a preselected glycan component which bears a fucosyl moiety, e.g., at a selected position on the glycan component, e.g., in a preparation of glycoproteins. 
         [0125]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. In another embodiment, the level of fucosylation is reduced by a predetermined level in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, cultured under reference conditions but otherwise the same or essentially the same as the cell cultured under conditions that result in said level of GDP-fucose. In another embodiment, the level of fucosylation is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0126]    In one embodiment, wherein X F  is greater than X G , 
         [0127]    and wherein, 
         [0128]    X F  is the % or proportion of reduction in the level of fucosylation (e.g., as compared to the level of fucosylation in a cell or batch of cultured cells cultured under reference conditions); and 
         [0129]    X G  is the % or proportion of reduction in the level of GDP fucose (as compared to the level of GDP fucose in a cell or batch of cultured cells cultured under reference conditions). 
         [0130]    In one embodiment, the cell or batch of cultured cells does not include an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0131]    In one embodiment, the cell or batch of cultured cells does includes an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0132]    In an embodiments, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. 
         [0133]    In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. 
         [0134]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. 
         [0135]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0136]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0137]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0138]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0139]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0140]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0141]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. 
         [0142]    In one embodiment, the method further comprises providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose. 
         [0000]    In another embodiment, the method further comprises providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below the second level. In another embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In one embodiment, the compound other than GDP-fucose is GDP-mannose. In one embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0143]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In one embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0144]    Methods described herein allow the production of proteins having reduced fucosylation from a cell line that is not genetically altered to reduce fucosylation. Such methods allow the use of a cell line that produces a reference glycoprotein, e.g., an approved product, by culturing that cell line to provide the reference glycoprotein with optimized levels of fucosylation. E.g., a cell line that has been optimized or otherwise selected for use in producing a protein, e.g., an FDA approved therapeutic protein, can be used to produce a protein having reduced fucosylation according to the invention, without genetically engineering the production line cell. 
         [0145]    Accordingly, in another aspect, the invention features, a method of providing a glycoprotein (or preparation thereof) having fucosylation that is reduced compared to a reference glycoprotein, e.g., an FDA approved glycoprotein. The method comprises: 
         [0146]    providing a cell that expresses said reference glycoprotein, which optionally, is wild-type for one or more (or all) of GMD, FX, fucokinase, GFPP, GDP-Fucose synthetase, a fucosyltransferase or a GDP-Fucose transporter; 
         [0147]    culturing said cell (without inducing a mutation in, or adding an siRNA that targets one or more of GMD, FX, fucokinase, GFPP, GDP-Fuc synthetase, a fucosyltransferase or a GDP-Fucose transporter) under culture conditions that result in a level of GDP-fucose in said cell that is below a first preselected level and, in embodiments, above a second preselected level, and results in a preselected level of fucosylation, which is less than in a reference cell cultured under reference conditions, e.g., to provide a batch of cultured cells; 
         [0148]    optionally, measuring the level of GDP-fucose in said cell or batch of cultured cells; and 
         [0149]    optionally, separating the glycoprotein from at least one component with which said cell or batch of cultured cells was cultured; 
         [0150]    optionally, evaluating the glycoprotein (or a glycoprotein on the surface of the cell or batch of cultured cells) for a parameter related to fucosylation; 
         [0151]    thereby providing a glycoprotein having fucosylation that is reduced compared to a reference glycoprotein, e.g., an FDA approved glycoprotein. 
         [0152]    In one embodiment, the method further comprises evaluating a glycan on the surface of said cell or batch of cultured cells in order to determine if the glycoprotein produced by said cell or batch of cultured cells has reduced fucosylation. In another embodiment, said evaluation comprises evaluating a glycan on the surface of said cell or batch of cultured cells, to determine a property of said glycan, comparing the property to a reference, to thereby determine if said glycan structure is present on the product. 
         [0153]    In one embodiment, the method further comprises evaluating a glycan on the surface of said cell or batch of cultured cells in order to determine if the glycoprotein produced by said cell or batch of cultured cells has reduced fucosylation. In another embodiment, said evaluation comprises evaluating a glycan on the surface of said cell or batch of cultured cells, to determine a property of said glycan, comparing the property to a reference, to thereby determine if said glycan structure is present on the product. 
         [0154]    In one embodiment, said first preselected level of GDP-fucose is selected from a level that is: 
         [0155]    i.a) approximately equal to or less than 80%, 70% or 60% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0156]    ii.a) approximately equal to, or less than, the point of maximum curvature above the inflection point (e.g., the inflection point in the second phase) on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0157]    ii.1.a) approximately equal to, or less than, the lowest level that results in a normal (e.g., that seen in an un-manipuated cell) level of fucosylation; 
         [0158]    iii.a) approximately equal to or less than the point of maximum curvature below the inflection point on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0159]    iii.1.a) approximately equal to, or less than, the highest level that results in no further reduction in fucosylation; 
         [0160]    iv.a) approximately equal to or less than point A on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; 
         [0161]    v.a) approximately equal to or less than that corresponding to an amount between points A and B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; or 
         [0162]    vi.a) approximately equal to or less than point B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0163]    In one embodiment, said second preselected level of GDP-fucose is selected from a level: 
         [0164]    i.b) approximately equal to, or greater than, 10%, 15%, 20%, 25%, 30%, 35% or 40% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0165]    ii.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in decrease of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, greater than, 10%, 20%, 30%, 40% or 50% than a reference levee, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0166]    iii.b) an amount that provides an unacceptable level of fucose deprivation, e.g. an amount that results in a level of high mannose structures that are less than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of a reference level; 
         [0167]    iv.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of GDP-mannose, e.g. an increase in GDP-mannose that is equal to or greater than 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level, e.g. the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0168]    v.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of high mannose structures that are more than or equal to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level; or 
         [0169]    vi.b) approximately equal to or greater than point C on the curve in  FIG. 1 , or greater than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0170]    In an embodiment the first level is i.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0171]    In an embodiment the first level is ii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0172]    In an embodiment the first level is ii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0173]    In an embodiment the first level is iii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0174]    In an embodiment the first level is iii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0175]    In an embodiment the first level is iv.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0176]    In an embodiment the first level is v.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0177]    In an embodiment the first level is vi.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0178]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is i.b. 
         [0179]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is ii.b. 
         [0180]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iii.b. 
         [0181]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iv.b. 
         [0182]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is v.b. 
         [0183]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is vi.b. 
         [0184]    In one embodiment, the level of GDP-fucose is selected to be outside the range between A and B on the curve in  FIG. 1  (as relatively small changes in GDP-fucose will result in relatively large changes in the amount of fucosylation. In an embodiment the level is also less than B.). In another embodiment, the level of GDP-fucose is reduced by a predetermined level, e.g., in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, cultured under reference conditions but otherwise the same or essentially the same as the cell cultured under conditions that result in said level of GDP-fucose. In another embodiment, the level of GDP-fucose is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0185]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the amount of fucosylation in the glycan complement, the amount or fucosylation on a component of the glycan complement, or the amount of fucosylation on a glycan component, e.g., in a preparation of glycoproteins. 
         [0186]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the proportion of a preselected glycan component which bears a fucosyl moiety, e.g., at a selected position on the glycan component, e.g., in a preparation of glycoproteins. 
         [0000]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. In another embodiment, the level of fucosylation is reduced by a predetermined level in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e e.g., a CHO cell or batch of cultured cells, cultured under reference conditions but otherwise the same or essentially the same as the cell cultured under conditions that result in said level of GDP-fucose. In another embodiment, the level of fucosylation is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0187]    In one embodiment, wherein X F  is greater than X G , 
         [0188]    and wherein, 
         [0189]    X F  is the % or proportion of reduction in the level of fucosylation (e.g., as compared to the level of fucosylation in a cell or batch of cultured cells cultured under reference conditions); and 
         [0190]    X G  is the % or proportion of reduction in the level of GDP fucose (as compared to the level of GDP fucose in a cell or batch of cultured cells cultured under reference conditions). 
         [0191]    In an embodiments, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. 
         [0192]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. 
         [0193]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0194]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0195]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0196]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0197]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0198]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0199]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. 
         [0200]    In one embodiment, the method further comprises providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose. 
         [0000]    In another embodiment, the method further comprises providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below the second level. In another embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In one embodiment, the compound other than GDP-fucose is GDP-mannose. In one embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0201]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In one embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0202]    In another aspect, the invention features, a reaction mixture containing one or more of a cell or batch of cultured cells having a manipulation, culture medium, and a glycoprotein having reduced fucosylation produced by the cell. 
         [0203]    In another aspect, the invention features, a device for the culture of cells comprising one or more of a cell having a manipulation, culture medium, and a glycoprotein having reduced fucosylation produced by the cell. 
         [0204]    When reduced fucosylation is desired, methods described herein allow selecting a cell which makes a desired protein, selecting a manipulation(s) that gives reduced fucosylation according to the invention, providing the manipulations to a cell, and optionally, using the cell for making the protein. Although useful in other applications, this method can be used to use and/or further modify an existing cell line that has been used to make a protein not having reduced fucosylation. 
         [0205]    Accordingly, in another aspect, the invention features, a method of making, or providing, a glycoprotein, or preparation thereof, having a glycan structure having reduced fucosylation, comprising: 
         [0206]    optionally, selecting a glycan structure having reduced fucosylation, e.g., from a list comprising a plurality of glycan structures having reduced fucosylation (in embodiments the list is provided), and optionally memorializing said selected glycan structure; 
         [0207]    selecting a cell, preferably on the basis that it produces a protein having the primary amino acid sequence of said glycoprotein but which protein lacks said glycan structure having reduced fucosylation; 
         [0208]    optionally, selecting a manipulation, e.g., selecting the manipulation on the basis that the manipulation decreases fucosylation and which manipulation thereby promotes the formation of said glycan structure having reduced fucosylation (in embodiments the manipulation is from a list comprising a plurality of manipulations, and in embodiments the list is provided); 
         [0209]    providing said manipulation to said cell to provide a cell having or subject to a manipulation that decreases the level of fucosylation and which manipulation thereby promotes the formation of said glycan structure having reduced fucosylation; 
         [0210]    culturing said selected cell, e.g., to provide a batch of cultured cells; 
         [0211]    optionally, separating the glycoprotein having a glycan structure from at least one component with which the cell or batch of cultured cells was cultured; 
         [0212]    optionally, analyzing said glycoprotein to confirm the presence of the glycan structure having reduced fucosylation; 
         [0213]    thereby making, or providing, a glycoprotein having a glycan structure having reduced fucosylation, e.g., by inhibiting or promoting the addition of a fucose moiety to a protein or glycoprotein. 
         [0214]    In one embodiment, the method further comprises evaluating a glycan on the surface of said cell or batch of cultured cells in order to determine if the glycoprotein produced by said cell or batch of cultured cells has reduced fucosylation. In another embodiment, said evaluation comprises evaluating a glycan on the surface of said cell or batch of cultured cells, to determine a property of said glycan, comparing the property to a reference, to thereby determine if said glycan structure is present on the product. 
         [0215]    In one embodiment, the manipulation results in a level of GDP-fucose in said cell that is below a first preselected level and, in embodiments, above a second preselected level. In embodiment said first preselected level of GDP-fucose is selected from a level that is: 
         [0216]    i.a) approximately equal to or less than 80%, 70% or 60% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0217]    ii.a) approximately equal to, or less than, the point of maximum curvature above the inflection point (e.g., the inflection point in the second phase) on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0218]    ii.1.a) approximately equal to, or less than, the lowest level that results in a normal (e.g., that seen in an un-manipuated cell) level of fucosylation; 
         [0219]    iii.a) approximately equal to or less than the point of maximum curvature below the inflection point on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0220]    iii.1.a) approximately equal to, or less than, the highest level that results in no further reduction in fucosylation; 
         [0221]    iv.a) approximately equal to or less than point A on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; 
         [0222]    v.a) approximately equal to or less than that corresponding to an amount between points A and B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; or 
         [0223]    vi.a) approximately equal to or less than point B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0224]    In one embodiment, said second preselected level of GDP-fucose is selected from a level: 
         [0225]    i.b) approximately equal to, or greater than, 10%, 15%, 20%, 25%, 30%, 35% or 40% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0226]    ii.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in decrease of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, greater than, 10%, 20%, 30%, 40% or 50% than a reference levee, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0227]    iii.b) an amount that provides an unacceptable level of fucose deprivation, e.g. an amount that results in a level of high mannose structures that are less than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of a reference level; 
         [0228]    iv.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of GDP-mannose, e.g. an increase in GDP-mannose that is equal to or greater than 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level, e.g. the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0229]    v.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of high mannose structures that are more than or equal to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level; or 
         [0230]    vi.b) approximately equal to or greater than point C on the curve in  FIG. 1 , or greater than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0231]    In an embodiment the first level is i.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0232]    In an embodiment the first level is ii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0233]    In an embodiment the first level is ii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0234]    In an embodiment the first level is iii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0235]    In an embodiment the first level is iii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0236]    In an embodiment the first level is iv.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0237]    In an embodiment the first level is v.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0238]    In an embodiment the first level is vi.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0239]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is i.b. 
         [0240]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is ii.b. 
         [0241]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iii.b. 
         [0242]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iv.b. 
         [0243]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is v.b. 
         [0244]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is vi.b. 
         [0245]    In an embodiment the level of GDP-fucose is between point B and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0246]    In an embodiment the level of GDP-fucose is between point A and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0247]    In one embodiment, the level of GDP-fucose is selected to be outside the range between A and B on the curve in  FIG. 1  (as relatively small changes in GDP-fucose will result in relatively large changes in the amount of fucosylation. In an embodiment the level is also less than B.) In another embodiment, the level of GDP-fucose is reduced by a predetermined level, e.g., in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell having the manipulation. In another embodiment, the level of GDP-fucose is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0248]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the amount of fucosylation in the glycan complement, the amount or fucosylation on a component of the glycan complement, or the amount of fucosylation on a glycan component, e.g., in a preparation of glycoproteins. 
         [0249]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the proportion of a preselected glycan component which bears a fucosyl moiety, e.g., at a selected position on the glycan component, e.g., in a preparation of glycoproteins. 
         [0250]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. In another embodiment, the level of fucosylation is reduced by a predetermined level in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell or batch of cultured cells having the manipulation. In another embodiment, the level of fucosylation is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0251]    In one embodiment, X F  is greater than X G , 
         [0252]    and wherein, 
         [0253]    X F  is the % or proportion of reduction in the level of fucosylation (e.g., as compared to the level of fucosylation in a cell or batch of cultured cells lacking the manipulation); and 
         [0254]    X G  is the % or proportion of reduction in the level of GDP fucose (as compared to the level of GDP fucose in a cell or batch of cultured cells lacking the manipulation). 
         [0255]    In one embodiment, said manipulation is not a genetic lesion or the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety. For example, the manipulation is not a lesion that decreases the expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells is wild-type for one or all of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells does not include an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, absent the manipulation, the level of fucosylation is substantially the same as the level in a wild-type cell. In another embodiment, the manipulated cell carries no mutation that substantially lowers GDP-fucose levels. In another embodiment, the manipulated cell has no siRNA that substantially lowers GDP-fucose levels. 
         [0256]    In one embodiment, the cell has a mutation (e.g., a genetically engineered change) that decreases the level of GDP-fucose. Exemplary mutations include those which alter the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. The mutation can be in the structural gene which encodes GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. Such mutations can decrease the activity of the encoded protein. The decrease can be partial or complete. Such mutations can act, e.g., by altering the catalytic activity of the protein or by altering its half-life. Other exemplary mutations can be in a sequence that control expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. These can be mutations that completely, or partially, reduce the expression of the gene, at the RNA or protein level. Such mutations include deletion or other mutations in endogenous of control sequence. Such mutations also include the introduction of heterologous control sequence, e.g., the introduction of heterologous control regions, e.g., a sequence that will give a desired level of expression. (A heterologous control sequence is a sequence other than a sequence naturally associated with and operably linked to the structural gene.) In embodiments the manipulation comprises a mutation in the structural region or in a control sequence operably linked to the gene. 
         [0257]    In an embodiment a cell having a mutation that that decreases the level of GDP-fucose, e.g., a mutation that decreases the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter is cultured in the presence of a substance, e.g., fucose, that results in a GDP-fucose level and/or a fucosylation level described herein. In an embodiment the cell includes a mutation that, in the absence of fucose in the culture medium, would result in a cell having an unacceptably low level of GDP-fucose. When, however, cultured under the appropriate conditions, e.g., media supplemented, e.g., with fucose, that cell can exhibit a desired level of GDP-fucose, e.g., a level of GDP-fucose described herein. Thus, fucose or another substance is present in the culture medium at a level that results in a level of GDP-fucose recited above. 
         [0258]    In another embodiment, the manipulation is the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety, e.g., an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter, and fucose or another substance is present in the culture medium at a level that results in formation of said glycoprotein having a glycan structure having reduced fucosylation. 
         [0259]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0260]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0261]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0262]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0263]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0264]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0265]    In one embodiment, the manipulation is, or is the product of, a selection for reduced levels of GDP-fucose. In another embodiment, the manipulation is, or is the product of, a selection for reduced fucosylation of a glycoprotein. In another embodiment, the manipulation comprises contact with, or inclusion in or on the cell or batch of cultured cells, of an exogenous inhibitor of an enzyme involved in GDP-fucose biosynthesis, e.g., a specific or non-specific inhibitor. 
         [0266]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. 
         [0267]    In one embodiment, one or more of said cell or said batch of cultured cells, said manipulation, and said glycoprotein, is selected on the basis that it or the combination will provide a glycoprotein having reduced fucosylation. 
         [0268]    In one embodiment, one or more of said cell or said batch of cultured cells, said manipulation (or manipulations), and said glycoprotein, is selected on the basis that it or the combination will provide a level of GDP-fucose described herein, e.g., a level which gives a minimal level of fucosylation (e.g., with reference to a curve analogous to that in  FIG. 1 , the level is to the right of point B) but which is above a preselected level. E.g., in a an embodiment the level is above a level that gives an unwanted decrease in the level of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, or more than, 10%, 20%, 30%, 40% or 50% as compared to a reference level, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation. 
         [0269]    In some embodiments the level is above a level that gives an unwanted increase in the level of GDP-mannose, e.g., an increase in GDP-mannose that is equal to, or more than, about 2×, 3×, 4×, 5×, ×, 7×, 8×, 9×, or 10× of a reference level, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation. 
         [0270]    In one embodiment, the method further comprises providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose. 
         [0000]    In another embodiment, the method further comprises providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below the second level. In another embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In one embodiment, the compound other than GDP-fucose is GDP-mannose. In one embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0271]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In one embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0272]    In an embodiments, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. The inhibitor can be used with a cell which is mutant or wildtype for one or more GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0273]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. Such media can be used, e.g., with a cell having a mutation that eliminates or decreased the activity of one or more of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0274]    When reduced fucosylation is desired, methods described herein allow selecting a cell which makes the desired protein. Although useful in other applications, this method can be used to use and/or further modify an existing cell line that has been used to make a protein not having reduced fucosylation. 
         [0275]    In one aspect, the invention features a method of providing a cell that makes a glycoprotein having a glycan structure having reduced fucosylation, comprising: 
         [0276]    optionally, selecting a glycan structure having reduced fucosylation, e.g., from a list comprising a plurality of glycan structures having reduced fucosylation (in embodiments the list is provided), and optionally memorializing said selected glycan structure; 
         [0277]    selecting a cell, preferably on the basis that it produces a protein having the primary amino acid sequence of said glycoprotein but which protein lacks said glycan structure having reduced fucosylation; 
         [0278]    optionally, selecting a manipulation, e.g., selecting the manipulation on the basis that the manipulation decreases the level of fucosylation, and which manipulation thereby promotes the formation of said glycan structure having reduced fucosylation (in embodiments the manipulation is from a list comprising a plurality of manipulations, and in embodiments the list is provided); 
         [0279]    providing said manipulation to said cell to provide a cell having or subject to a manipulation that decreases fucosylation, and which manipulation thereby promotes the formation of said glycan structure having reduced fucosylation; 
         [0280]    optionally producing glycoprotein from said cell and determining if said glycoprotein has said glycan structure having reduced fucosylation, thereby providing a cell that makes a glycoprotein having a glycan structure. 
         [0281]    In one embodiment, the method further comprises evaluating a glycan on the surface of said cell or batch of cultured cells in order to determine if the glycoprotein produced by said cell or batch of cultured cells has reduced fucosylation. In another embodiment, said evaluation comprises evaluating a glycan on the surface of said cell or batch of cultured cells, to determine a property of said glycan, comparing the property to a reference, to thereby determine if said glycan structure is present on the product. 
         [0282]    In one embodiment, the manipulation results in a level of GDP-fucose in said cell that is below a first preselected level and, in embodiments, above a second preselected level. In one embodiment, said first preselected level of GDP-fucose is selected from a level that is: 
         [0283]    i.a) approximately equal to or less than 80%, 70% or 60% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0284]    ii.a) approximately equal to, or less than, the point of maximum curvature above the inflection point (e.g., the inflection point in the second phase) on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0285]    ii.1.a) approximately equal to, or less than, the lowest level that results in a normal (e.g., that seen in an un-manipuated cell) level of fucosylation; 
         [0286]    iii.a) approximately equal to or less than the point of maximum curvature below the inflection point on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0287]    iii.1.a) approximately equal to, or less than, the highest level that results in no further reduction in fucosylation; 
         [0288]    iv.a) approximately equal to or less than point A on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; 
         [0289]    v.a) approximately equal to or less than that corresponding to an amount between points A and B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; or 
         [0290]    vi.a) approximately equal to or less than point B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0291]    In one embodiment, said second preselected level of GDP-fucose is selected from a level: 
         [0292]    i.b) approximately equal to, or greater than, 10%, 15%, 20%, 25%, 30%, 35% or 40% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0293]    ii.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in decrease of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, greater than, 10%, 20%, 30%, 40% or 50% than a reference levee, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0294]    iii.b) an amount that provides an unacceptable level of fucose deprivation, e.g. an amount that results in a level of high mannose structures that are less than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of a reference level; 
         [0295]    iv.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of GDP-mannose, e.g. an increase in GDP-mannose that is equal to or greater than 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level, e.g. the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0296]    v.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of high mannose structures that are more than or equal to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level; or 
         [0297]    vi.b) approximately equal to or greater than point C on the curve in  FIG. 1 , or greater than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0298]    In an embodiment the first level is i.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0299]    In an embodiment the first level is ii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0300]    In an embodiment the first level is ii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0301]    In an embodiment the first level is iii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0302]    In an embodiment the first level is iii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0303]    In an embodiment the first level is iv.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0304]    In an embodiment the first level is v.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0305]    In an embodiment the first level is vi.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0306]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is i.b. 
         [0307]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is ii.b. 
         [0308]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iii.b. 
         [0309]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iv.b. 
         [0310]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is v.b. 
         [0311]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is vi.b. 
         [0312]    In an embodiment the level of GDP-fucose is between point B and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0313]    In an embodiment the level of GDP-fucose is between point A and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0314]    In one embodiment, the level of GDP-fucose is selected to be outside the range between A and B on the curve in  FIG. 1  (as relatively small changes in GDP-fucose will result in relatively large changes in the amount of fucosylation. In an embodiment the level is also less than B.) In another embodiment, the level of GDP-fucose is reduced by a predetermined level, e.g., in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell having the manipulation. In another embodiment, the level of GDP-fucose is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0000]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the amount of fucosylation in the glycan complement, the amount or fucosylation on a component of the glycan complement, or the amount of fucosylation on a glycan component, e.g., in a preparation of glycoproteins. 
         [0315]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the proportion of a preselected glycan component which bears a fucosyl moiety, e.g., at a selected position on the glycan component, e.g., in a preparation of glycoproteins. 
         [0316]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. In another embodiment, the level of fucosylation is reduced by a predetermined level in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell or batch of cultured cells having the manipulation. In another embodiment, the level of fucosylation is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0317]    In one embodiment, X F  is greater than X G , 
         [0318]    and wherein, 
         [0319]    X F  is the % or proportion of reduction in the level of fucosylation (e.g., as compared to the level of fucosylation in a cell or batch of cultured cells lacking the manipulation); and 
         [0320]    X G  is the % or proportion of reduction in the level of GDP fucose (as compared to the level of GDP fucose in a cell or batch of cultured cells lacking the manipulation). 
         [0321]    In one embodiment, said manipulation is not a genetic lesion or the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety. For example, the manipulation is not a lesion that decreases the expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells is wild-type for one or all of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells does not include an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, absent the manipulation, the level of fucosylation is substantially the same as the level in a wild-type cell. In another embodiment, the manipulated cell carries no mutation that substantially lowers GDP-fucose levels. In another embodiment, the manipulated cell has no siRNA that substantially lowers GDP-fucose levels. 
         [0322]    In one embodiment, the cell has a mutation (e.g., a genetically engineered change) that decreases the level of GDP-fucose. Exemplary mutations include those which alter the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0000]    The mutation can be in the structural gene which encodes GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. Such mutations can decrease the activity of the encoded protein. The decrease can be partial or complete. Such mutations can act, e.g., by altering the catalytic activity of the protein or by altering its half-life. Other exemplary mutations can be in a sequence that control expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. These can be mutations that completely, or partially, reduce the expression of the gene, at the RNA or protein level. Such mutations include deletion or other mutations in endogenous of control sequence. Such mutations also include the introduction of heterologous control sequence, e.g., the introduction of heterologous control regions, e.g., a sequence that will give a desired level of expression. (A heterologous control sequence is a sequence other than a sequence naturally associated with and operably linked to the structural gene.) In embodiments the manipulation comprises a mutation in the structural region or in a control sequence operably linked to the gene. 
         [0323]    In an embodiment a cell having a mutation that that decreases the level of GDP-fucose, e.g., a mutation that decreases the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter is cultured in the presence of a substance, e.g., fucose, that results in a GDP-fucose level and/or a fucosylation level described herein. In an embodiment the cell includes a mutation that, in the absence of fucose in the culture medium, would result in a cell having an unacceptably low level of GDP-fucose. When, however, cultured under the appropriate conditions, e.g., media supplemented, e.g., with fucose, that cell can exhibit a desired level of GDP-fucose, e.g., a level of GDP-fucose described herein. Thus, fucose or another substance is present in the culture medium at a level that results in a level of GDP-fucose recited above. 
         [0324]    In another embodiment, the manipulation is the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety, e.g., an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter, and fucose or another substance is present in the culture medium at a level that results in formation of said glycoprotein having a glycan structure having reduced fucosylation. 
         [0325]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0326]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0327]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0328]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0329]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0330]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0331]    In one embodiment, the manipulation is, or is the product of, a selection for reduced levels of GDP-fucose. In another embodiment, the manipulation is, or is the product of, a selection for reduced fucosylation of a glycoprotein. In another embodiment, the manipulation comprises contact with, or inclusion in or on the cell or batch of cultured cells, of an exogenous inhibitor of an enzyme involved in GDP-fucose biosynthesis, e.g., a specific or non-specific inhibitor. 
         [0332]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. 
         [0333]    In one embodiment, one or more of said cell or said batch of cultured cells, said manipulation, and said glycoprotein, is selected on the basis that it or the combination will provide a glycoprotein having reduced fucosylation. 
         [0334]    In one embodiment, one or more of said cell or said batch of cultured cells, said manipulation (or manipulations), and said glycoprotein, is selected on the basis that it or the combination will provide a level of GDP-fucose described herein, e.g., a level which gives a minimal level of fucosylation (e.g., with reference to a curve analogous to that in  FIG. 1 , the level is to the right of point B) but which is above a preselected level, e.g., above a level that gives an unwanted decrease in the level of GDP-mannose. E.g., the level is above a level that gives a decrease in GDP-mannose that is equal to, or more than, 10%, 20%, 30%, 40% or 50% as compared to a reference level, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation. 
         [0335]    In some embodiments the level is above a level that gives an unwanted increase in the level of GDP-mannose, e.g., an increase in GDP-mannose that is equal to, or more than, about 2×, 3×, 4×, 5×, ×, 7×, 8×, 9×, or 10× of a reference level, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation. 
         [0336]    In one embodiment, the method further comprises providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose. 
         [0000]    In another embodiment, the method further comprises providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below the second level. In another embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In one embodiment, the compound other than GDP-fucose is GDP-mannose. In one embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0337]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In one embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0338]    In an embodiments, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. The inhibitor can be used with a cell which is mutant or wildtype for one or more GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0339]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. Such media can be used, e.g., with a cell having a mutation that eliminates or decreased the activity of one or more of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0340]    Methods described herein allow monitoring a process of making a protein, e.g., to insure that the process is in compliance with parameters set out herein. 
         [0341]    Thus, in another aspect, the invention features, a method of monitoring a process, e.g., a process of culturing cells, e.g., of a selected type, to produce a product, comprising: 
         [0342]    optionally, selecting a glycan structure having reduced fucosylation, e.g., from a list comprising a plurality of glycan structures having reduced fucosylation (in embodiments the list is provided), and optionally memorializing said selected glycan structure; 
         [0343]    optionally, selecting a cell on the basis of the cell having or subject to a manipulation that decreases the level of fucosylation or GDP-fucose, and which manipulation decreases the level of fucosylation or GDP-fucose (in embodiments the manipulation is from a list comprising a plurality of manipulations, and in embodiments the list is provided); 
         [0344]    providing a cell having or subject to a manipulation that decreases the level of fucosylation or GDP-fucose, e.g., a cell having a manipulation described herein or a cell a cell selected by a method described herein; 
         [0345]    culturing said cell, e.g., to provide a batch of cultured cells; and 
         [0346]    evaluating (directly or indirectly) the level of GDP-fucose of, or a glycan complement, glycan component or glycan structure produced by, the cell or the batch of cultured cells, 
         [0000]    to thereby monitor the process. 
         [0347]    In one embodiment, the evaluating step comprises any of: 
         [0348]    (a) isolating glycoproteins produced from the cell or the batch of cultured cells and evaluating the glycans containing on the glycoproteins, 
         [0349]    (b) isolating a specific glycoprotein composition produced from the cell or the batch of cultured cells and evaluating the glycans from the isolated glycoprotein composition, 
         [0350]    (c) obtaining a glycan preparation from a glycoprotein preparation or isolated glycoprotein produced from the cell or the batch of cultured cells and evaluating the glycans in the glycan preparation, 
         [0351]    (d) cleaving monosaccharides from glycans present on a glycoprotein produced from the cell or the batch of cultured cells or from glycans on the surface of the cell or the batch of cultured cells, and detecting the cleaved monosaccharides, 
         [0352]    (e) providing at least one peptide from a glycoprotein preparation produced from the cell or the batch of cultured cells, and evaluating the glycans on the at least one peptide, and 
         [0353]    (f) evaluating glycans from glycans on the cell surface of the cell or the batch of cultured cells. 
         [0354]    In another embodiment, the evaluating step comprises isolating glycoproteins produced from the cell or the batch of cultured cells and evaluating the glycans containing on the glycoproteins. In another embodiment, the evaluating step comprises isolating a specific glycoprotein composition produced from the cell or the batch of cultured cells and evaluating the glycans from the isolated glycoprotein composition. In another embodiment, the evaluating step comprises obtaining a glycan preparation from a glycoprotein preparation or isolated glycoprotein produced from the cell or the batch of cultured cells and evaluating the glycans in the glycan preparation. In another embodiment, the evaluating step comprises cleaving monosaccharides from glycans present on a glycoprotein produced from the cell or the batch of cultured cells or from glycans on the surface of the cell or the batch of cultured cells, and detecting the cleaved monosaccharides. In another embodiment, the evaluating step comprises providing at least one peptide from a glycoprotein preparation produced from the cell or the batch of cultured cells, and evaluating the glycans on the at least one peptide. In another embodiment, the evaluating step comprises evaluating glycans from glycans on the cell surface of the cell or the batch of cultured cells. 
         [0355]    In another embodiment, the method further comprises, if an observed value from an evaluation step does not meet a reference value, discarding said cell, continuing culture of said cell, or altering a culture condition and further culturing said cell. In another embodiment, the method further comprises, if an observed value from an evaluation step meets said reference value, continuing culture of said cell or said batch of cultured cells, altering a culture condition and further culturing said cell or said batch of cultured cells, or discarding said cell or said batch of cultured cells. In another embodiment, the method further comprises continuing culture of the cell or the batch of cultured cells. In another embodiment, the method further comprises altering a culture condition and further culturing said cell or said batch of cultured cells and optionally repeating the evaluation. 
         [0356]    In one embodiment, the evaluation comprises determining if the level of GDP-fucose in said cell that is below a first preselected level and, in embodiments, above a second preselected level. In one embodiment, said first preselected level of GDP-fucose is selected from a level that is: 
         [0357]    i.a) approximately equal to or less than 80%, 70% or 60% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0358]    ii.a) approximately equal to, or less than, the point of maximum curvature above the inflection point (e.g., the inflection point in the second phase) on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0359]    ii.1.a) approximately equal to, or less than, the lowest level that results in a normal (e.g., that seen in an un-manipuated cell) level of fucosylation; 
         [0360]    iii.a) approximately equal to or less than the point of maximum curvature below the inflection point on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0361]    iii.1.a) approximately equal to, or less than, the highest level that results in no further reduction in fucosylation; 
         [0362]    iv.a) approximately equal to or less than point A on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; 
         [0363]    v.a) approximately equal to or less than that corresponding to an amount between points A and B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; or 
         [0364]    vi.a) approximately equal to or less than point B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0365]    In one embodiment, said second preselected level of GDP-fucose is selected from a level: 
         [0366]    i.b) approximately equal to, or greater than, 10%, 15%, 20%, 25%, 30%, 35% or 40% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0367]    ii.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in decrease of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, greater than, 10%, 20%, 30%, 40% or 50% than a reference levee, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0368]    iii.b) an amount that provides an unacceptable level of fucose deprivation, e.g. an amount that results in a level of high mannose structures that are less than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of a reference level; 
         [0369]    iv.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of GDP-mannose, e.g. an increase in GDP-mannose that is equal to or greater than 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level, e.g. the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0370]    v.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of high mannose structures that are more than or equal to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level; or 
         [0371]    vi.b) approximately equal to or greater than point C on the curve in  FIG. 1 , or greater than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0372]    In an embodiment the first level is i.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0373]    In an embodiment the first level is ii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0374]    In an embodiment the first level is ii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0375]    In an embodiment the first level is iii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0376]    In an embodiment the first level is iii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0377]    In an embodiment the first level is iv.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0378]    In an embodiment the first level is v.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0379]    In an embodiment the first level is vi.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0380]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is i.b. 
         [0381]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is ii.b. 
         [0382]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iii.b. 
         [0383]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iv.b. 
         [0384]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is v.b. 
         [0385]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is vi.b. 
         [0386]    In an embodiment the level of GDP-fucose is between point B and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0387]    In an embodiment the level of GDP-fucose is between point A and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0388]    In one embodiment, the level of GDP-fucose is selected to be outside the range between A and B on the curve in  FIG. 1  (as relatively small changes in GDP-fucose will result in relatively large changes in the amount of fucosylation. In an embodiment the level is also less than B.) In another embodiment, the level of GDP-fucose is reduced by a predetermined level, e.g., in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell having the manipulation. In another embodiment, the level of GDP-fucose is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0389]    In one embodiment, said manipulation is not a genetic lesion or the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety. For example, the manipulation is not a lesion that decreases the expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells is wild-type for one or all of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells does not include an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, absent the manipulation, the level of fucosylation is substantially the same as the level in a wild-type cell. In another embodiment, the manipulated cell carries no mutation that substantially lowers GDP-fucose levels. In another embodiment, the manipulated cell has no siRNA that substantially lowers GDP-fucose levels. 
         [0390]    In one embodiment, the cell has a mutation (e.g., a genetically engineered change) that decreases the level of GDP-fucose. Exemplary mutations include those which alter the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. The mutation can be in the structural gene which encodes GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. Such mutations can decrease the activity of the encoded protein. The decrease can be partial or complete. Such mutations can act, e.g., by altering the catalytic activity of the protein or by altering its half-life. Other exemplary mutations can be in a sequences that control expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. These can be mutations that completely, or partially, reduce the expression of the gene, at the RNA or protein level. Such mutations include deletion or other mutations in endogenous of control sequence. Such mutations also include the introduction of heterologous control sequence, e.g., the introduction of heterologous control regions, e.g., a sequence that will give a desired level of expression. (A heterologous control sequence is a sequence other than a sequence naturally associated with and operably linked to the structural gene.) In embodiments the manipulation comprises a mutation in the structural region or in a control sequence operably linked to the gene. 
         [0391]    In an embodiment a cell having a mutation that that decreases the level of GDP-fucose, e.g., a mutation that decreases the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter is cultured in the presence of a substance, e.g., fucose, that results in a GDP-fucose level and/or a fucosylation level described herein. In an embodiment the cell includes a mutation that, in the absence of fucose in the culture medium, would result in a cell having an unacceptably low level of GDP-fucose. When, however, cultured under the appropriate conditions, e.g., media supplemented, e.g., with fucose, that cell can exhibit a desired level of GDP-fucose, e.g., a level of GDP-fucose described herein. Thus, fucose or another substance is present in the culture medium at a level that results in a level of GDP-fucose recited above. 
         [0392]    In another embodiment, the manipulation is the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety, e.g., an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter, and fucose or another substance is present in the culture medium at a level that results in formation of said glycan structure having reduced fucosylation. 
         [0393]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0394]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0395]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0396]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0397]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0398]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0399]    In one embodiment, the manipulation is, or is the product of, a selection for reduced levels of GDP-fucose. In another embodiment, the manipulation is, or is the product of, a selection for reduced fucosylation of a glycoprotein. In another embodiment, the manipulation comprises contact with, or inclusion in or on the cell or batch of cultured cells, of an exogenous inhibitor of an enzyme involved in GDP-fucose biosynthesis, e.g., a specific or non-specific inhibitor. 
         [0400]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. 
         [0401]    In one embodiment, the method further comprises providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose. 
         [0000]    In another embodiment, the method further comprises providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below the second level. In another embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In one embodiment, the compound other than GDP-fucose is GDP-mannose. In one embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0402]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In one embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0403]    In an embodiments, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. The inhibitor can be used with a cell which is mutant or wildtype for one or more GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0404]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. Such media can be used, e.g., with a cell having a mutation that eliminates or decreased the activity of one or more of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0405]    Methods described herein allow monitoring a process of making a protein, e.g., to insure that the process is in compliance with parameters set out herein. 
         [0406]    In one aspect, the invention features a method of controlling a process for making a glycoprotein having a glycan structure with reduced fucosylation, comprising:
       (1) providing a glycoprotein made by the process of       
 
         [0408]    optionally, selecting a glycan structure having reduced fucosylation, e.g., from a list comprising a plurality of glycan structures having reduced fucosylation (in embodiments the list is provided); 
         [0409]    optionally, selecting a cell on the basis of the cell having or subject to a manipulation that decreases the level of fucosylation or GDP-fucose, and which manipulation decreases the level of fucosylation or GDP-fucose (in embodiments the manipulation is from a list comprising a plurality of manipulations, and in embodiments the list is provided); 
         [0410]    providing a cell having or subject to a manipulation that decreases the level of decreases the level of fucosylation or GDP-fucose; and 
         [0411]    culturing the cell to provide a glycoprotein and, e.g., form a batch of cultured cells;
       (2) evaluating (directly or indirectly) the level of GDP-fucose in the cells or the glycan structure of the glycoprotein,   (3) responsive to said evaluation, selecting a production parameter, e.g., a culture
 
condition, e.g., a level of a nutrient or other component in the culture medium, e.g., to provide a selected level of GDP-fucose in the cells or the selected glycan structure of the glycoprotein,
       
 
         [0414]    to thereby control the process for making a glycoprotein having a glycan structure. 
         [0415]    In one embodiment, the method comprises continuing culture of the cell or batch of cultured cells under conditions that differ from those used prior to the evaluation. In another embodiment, the method comprises continuing culture of the cell or batch of cultured cells under the same conditions used prior to the evaluation. 
         [0416]    In one embodiment, the evaluation comprises determining if the level of GDP-fucose in said cell that is below a first preselected level and, in embodiments, above a second preselected level. In one embodiment, said first preselected level of GDP-fucose is selected from a level that is: 
         [0417]    i.a) approximately equal to or less than 80%, 70% or 60% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0418]    ii.a) approximately equal to, or less than, the point of maximum curvature above the inflection point (e.g., the inflection point in the second phase) on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0419]    ii.1.a) approximately equal to, or less than, the lowest level that results in a normal (e.g., that seen in an un-manipuated cell) level of fucosylation; 
         [0420]    iii.a) approximately equal to or less than the point of maximum curvature below the inflection point on a graph of the amount of fucosylation vs. decrease in GDP-fucose; 
         [0421]    iii.1.a) approximately equal to, or less than, the highest level that results in no further reduction in fucosylation; 
         [0422]    iv.a) approximately equal to or less than point A on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; 
         [0423]    v.a) approximately equal to or less than that corresponding to an amount between points A and B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control; or 
         [0424]    vi.a) approximately equal to or less than point B on the curve in  FIG. 1 , or less than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0425]    In one embodiment, said second preselected level of GDP-fucose is selected from a level: 
         [0426]    i.b) approximately equal to, or greater than, 10%, 15%, 20%, 25%, 30%, 35% or 40% of a reference level, e.g., the level in said cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0427]    ii.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in decrease of GDP-mannose, e.g., a decrease in GDP-mannose that is equal to, greater than, 10%, 20%, 30%, 40% or 50% than a reference levee, e.g., the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0428]    iii.b) an amount that provides an unacceptable level of fucose deprivation, e.g. an amount that results in a level of high mannose structures that are less than or equal to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of a reference level; 
         [0429]    iv.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of GDP-mannose, e.g. an increase in GDP-mannose that is equal to or greater than 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level, e.g. the level of GDP-mannose in a cell or batch of cultured cells, e.g., a cell or batch of cultured cells which is otherwise similar, without the manipulation; 
         [0430]    v.b) an amount that provides an unacceptable level of fucose deprivation, e.g., an amount that results in accumulation of high mannose structures that are more than or equal to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, or 10× of a reference level; or 
         [0431]    vi.b) approximately equal to or greater than point C on the curve in  FIG. 1 , or greater than or equal to an analogous point on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0432]    In an embodiment the first level is i.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0433]    In an embodiment the first level is ii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0434]    In an embodiment the first level is ii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0435]    In an embodiment the first level is iii.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0436]    In an embodiment the first level is iii.1.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0437]    In an embodiment the first level is iv.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0438]    In an embodiment the first level is v.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0439]    In an embodiment the first level is vi.a and the second level is selected from i.b, ii.b, iii.b, iv.b, v.b, and vi.b. 
         [0440]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is i.b. 
         [0441]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is ii.b. 
         [0442]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iii.b. 
         [0443]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is iv.b. 
         [0444]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is v.b. 
         [0445]    In an embodiment the first level is selected from i.a, ii.a, ii.1.a, iii.a, iii.1.a, iv.a, v.a, and vi.a and the second level is vi.b. 
         [0446]    In an embodiment the level of GDP-fucose is between point B and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0447]    In an embodiment the level of GDP-fucose is between point A and C on the curve in  FIG. 1  or in an analogous range on a plot of the amount of fucosylation (%) vs. the amount of GDP fucose as a % of control. 
         [0448]    In one embodiment, the level of GDP-fucose is selected to be outside the range between A and B on the curve in  FIG. 1  (as relatively small changes in GDP-fucose will result in relatively large changes in the amount of fucosylation. In an embodiment the level is also less than B.) In another embodiment, the level of GDP-fucose is reduced by a predetermined level, e.g., in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell having the manipulation. In another embodiment, the level of GDP-fucose is reduced by, as much as, or more than, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0449]    In one embodiment, said evaluation step comprises comparing the structure of said glycan structure having reduced fucosylation present on a glycoprotein from said cultured cell or batch of cultured cells to a reference, and determining if said glycan structure having reduced fucosylation present on a glycoprotein from said cultured cell or batch of cultured cells differs from the corresponding glycan structure formed by a cell or batch of cultured cells that lacks the manipulation. 
         [0450]    In one embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the amount of fucosylation in the glycan complement, the amount or fucosylation on a component of the glycan complement, or the amount of fucosylation on a glycan component, e.g., in a preparation of glycoproteins. In another embodiment, the method further comprises evaluating the glycoprotein for a parameter related to fucosylation, e.g., the proportion of a preselected glycan component which bears a fucosyl moiety, e.g., at a selected position on the glycan component, e.g., in a preparation of glycoproteins. 
         [0451]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. In another embodiment, the level of fucosylation is reduced by a predetermined level in comparison with a reference. In another embodiment, the reference is the amount present in a cell or batch of cultured cells, e.g., a CHO cell or batch of cultured cells, lacking the manipulation but otherwise the same or essentially the same as the cell or batch of cultured cells having the manipulation. In another embodiment, the level of fucosylation is reduced by, as much as, or more than, 10, 20, 30, 40, 50, 60, 70, 80 or 90%, as compared to the reference. 
         [0452]    In one embodiment, said manipulation is not a genetic lesion or the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety. For example, the manipulation is not a lesion that decreases the expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells is wild-type for one or all of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells does not include an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, absent the manipulation, the level of fucosylation is substantially the same as the level in a wild-type cell. In another embodiment, the manipulated cell carries no mutation that substantially lowers GDP-fucose levels. In another embodiment, the manipulated cell has no siRNA that substantially lowers GDP-fucose levels. 
         [0453]    In one embodiment, the cell has a mutation (e.g., a genetically engineered change) that decreases the level of GDP-fucose. Exemplary mutations include those which alter the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0000]    The mutation can be in the structural gene which encodes GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. Such mutations can decrease the activity of the encoded protein. The decrease can be partial or complete. Such mutations can act, e.g., by altering the catalytic activity of the protein or by altering its half-life. Other exemplary mutations can be in a sequences that control expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. These can be mutations that completely, or partially, reduce the expression of the gene, at the RNA or protein level. Such mutations include deletion or other mutations in endogenous of control sequence. Such mutations also include the introduction of heterologous control sequence, e.g., the introduction of heterologous control regions, e.g., a sequence that will give a desired level of expression. (A heterologous control sequence is a sequence other than a sequence naturally associated with and operably linked to the structural gene.) In embodiments the manipulation comprises a mutation in the structural region or in a control sequence operably linked to the gene. 
         [0454]    In an embodiment a cell having a mutation that that decreases the level of GDP-fucose, e.g., a mutation that decreases the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter is cultured in the presence of a substance, e.g., fucose, that results in a GDP-fucose level and/or a fucosylation level described herein. In an embodiment the cell includes a mutation that, in the absence of fucose in the culture medium, would result in a cell having an unacceptably low level of GDP-fucose. When, however, cultured under the appropriate conditions, e.g., media supplemented, e.g., with fucose, that cell can exhibit a desired level of GDP-fucose, e.g., a level of GDP-fucose described herein. Thus, fucose or another substance is present in the culture medium at a level that results in a level of GDP-fucose recited above. 
         [0455]    In another embodiment, the manipulation is the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety, e.g., an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter, and fucose or another substance is present in the culture medium at a level that results in formation of said glycan structure having reduced fucosylation. 
         [0456]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0457]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0458]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0459]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0460]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0461]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0462]    In one embodiment, the manipulation is, or is the product of, a selection for reduced levels of GDP-fucose. In another embodiment, the manipulation is, or is the product of, a selection for reduced fucosylation of a glycoprotein. In another embodiment, the manipulation comprises contact with, or inclusion in or on the cell or batch of cultured cells, of an exogenous inhibitor of an enzyme involved in GDP-fucose biosynthesis, e.g., a specific or non-specific inhibitor. 
         [0463]    In one embodiment, the level of fucosylation at one, two, three, or more preselected amino acid residues is evaluated. 
         [0464]    In one embodiment, one or more of said cell or said batch of cultured cells, said manipulation, and said glycoprotein, is selected on the basis that it or the combination will provide a glycoprotein having reduced fucosylation. 
         [0465]    In one embodiment, the method further comprises providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose. 
         [0000]    In another embodiment, the method further comprises providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below the second level. In another embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In one embodiment, the compound other than GDP-fucose is GDP-mannose. In one embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0466]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In one embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0467]    In an embodiments, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. The inhibitor can be used with a cell which is mutant or wildtype for one or more GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0468]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. Such media can be used, e.g., with a cell having a mutation that eliminates or decreased the activity of one or more of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0469]    Methods described herein allow monitoring a process of making a protein, e.g., to insure that the process is in compliance with parameters set out herein. 
         [0470]    In one aspect, the invention features method of controlling a process for making a glycoprotein having a glycan structure with reduced fucosylation, comprising: 
         [0471]    (1) providing a glycoprotein made by the process of: 
         [0472]    optionally, selecting a glycan structure having reduced fucosylation, e.g., from a list comprising a plurality of glycan structures having reduced fucosylation (in embodiments the list is provided); 
         [0473]    optionally, selecting a cell on the basis of the cell having or subject to a manipulation that decreases the level of fucosylation or GDP-fucose, and which manipulation decreases the level of fucosylation or GDP-fucose (in embodiments the manipulation is from a list comprising a plurality of manipulations, and in embodiments the list is provided); 
         [0474]    providing a cell having or subject to a manipulation that decreases the level of decreases the level of fucosylation or GDP-fucose; and 
         [0475]    culturing the cell to provide a glycoprotein and, e.g., form a batch of cultured cells; 
         [0476]    (2) providing a value for a parameter associated with a compound other than GDP-fucose, wherein a parameter for the compound, e.g., the level of the compound, is correlated to the level of GDP-fucose, 
         [0477]    (3) providing a comparison of the value with a reference value, wherein optionally, a preselected relationship of the value to the reference value, e.g., greater than, equal to, or less than, is indicative of whether the level of GDP fucose is above, at or below a preselected level 
         [0478]    (4) responsive to said comparison, selecting a production parameter, e.g., a culture condition, e.g., a level of a nutrient or other component in the culture medium, to thereby control the process for making a glycoprotein having a glycan structure. 
         [0479]    In one embodiment, the method further comprises, responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture without intervening to change the level of GDP-fucose. In another embodiment, the compound other than GDP-fucose is GDP-mannose. In another embodiment, the compound other than GDP-fucose is GDP-mannose and the parameter is the level of GDP-mannose. 
         [0480]    In one embodiment, the method further comprises providing a value for the level of GDP-mannose, providing a comparison of the value with a reference value, and responsive to the result of the comparison, increasing the level of GDP-fucose, decreasing the level of GDP-fucose or continuing cell culture at without intervening to change the level of GDP-fucose. In another embodiment, the method comprises continuing to culture said cells, and repeating the steps above. 
         [0481]    In one embodiment, said manipulation is not a genetic lesion or the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety. For example, the manipulation is not a lesion that decreases the expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells is wild-type for one or all of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, the cell or batch of cultured cells does not include an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. In another embodiment, absent the manipulation, the level of fucosylation is substantially the same as the level in a wild-type cell. In another embodiment, the manipulated cell carries no mutation that substantially lowers GDP-fucose levels. In another embodiment, the manipulated cell has no siRNA that substantially lowers GDP-fucose levels. 
         [0482]    In one embodiment, the cell has a mutation (e.g., a genetically engineered change) that decreases the level of GDP-fucose. Exemplary mutations include those which alter the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. The mutation can be in the structural gene which encodes GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. Such mutations can decrease the activity of the encoded protein. The decrease can be partial or complete. Such mutations can act, e.g., by altering the catalytic activity of the protein or by altering its half-life. Other exemplary mutations can be in a sequences that control expression of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. These can be mutations that completely, or partially, reduce the expression of the gene, at the RNA or protein level. Such mutations include deletion or other mutations in endogenous of control sequence. Such mutations also include the introduction of heterologous control sequence, e.g., the introduction of heterologous control regions, e.g., a sequence that will give a desired level of expression. (A heterologous control sequence is a sequence other than a sequence naturally associated with and operably linked to the structural gene.) In embodiments the manipulation comprises a mutation in the structural region or in a control sequence operably linked to the gene. 
         [0483]    In an embodiment a cell having a mutation that that decreases the level of GDP-fucose, e.g., a mutation that decreases the activity of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter is cultured in the presence of a substance, e.g., fucose, that results in a GDP-fucose level and/or a fucosylation level described herein. In an embodiment the cell includes a mutation that, in the absence of fucose in the culture medium, would result in a cell having an unacceptably low level of GDP-fucose. When, however, cultured under the appropriate conditions, e.g., media supplemented, e.g., with fucose, that cell can exhibit a desired level of GDP-fucose, e.g., a level of GDP-fucose described herein. Thus, fucose or another substance is present in the culture medium at a level that results in a level of GDP-fucose recited above. 
         [0484]    In another embodiment, the manipulation is the presence of an siRNA that reduces the level of an enzyme that promotes formation of GDP-fucose, or the attachment of a fucosyl moiety, e.g., an siRNA that targets GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter, and fucose or another substance is present in the culture medium at a level that results in formation of said glycan structure having reduced fucosylation. 
         [0485]    In one embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0486]    In one embodiment, the cell is a Chinese Hamster Ovary (CHO) cell. In another embodiment, the glycoprotein is an antibody. In another embodiment, the antibody has reduced core fucosylation. In another embodiment, the antibody is selected from the group consisting of Rituximab, Trastuzamab, Bevacizumab, Tositumomab, Alemtuzumab, Arcitumomab, Cetuximab, Trastuzumab, Adalimumab, Ranibizumab, Gemtuzumab [ozogamicin], Fanolesomab, Efalizumab, Infliximab, Abciximab, Rituximab, Basiliximab, Eculizumab, Palivizumab, Natalizumab, Omalizumab, Daclizumab, and Ibritumomab. 
         [0487]    In one embodiment, the glycoprotein is selected from Table 1. 
         [0488]    In one embodiment, the method further comprises culturing a plurality of the cells and separating as much as, or at least, 1, 10, 100, 1,000, or 10,000 grams of the glycoprotein from the cells. In another embodiment, the method further comprises combining the glycoprotein having reduced fucosylation with a pharmaceutically acceptable component and, e.g., formulating the glycoprotein having reduced fucosylation into a pharmaceutically acceptable formulation. 
         [0489]    In one embodiment, the glycoprotein is analyzed by one or more of HPLC, CE, MALDI-MS and NMR. 
         [0490]    In one embodiment, the method further comprises memorializing the result of the evaluation. 
         [0491]    In one embodiment, the manipulation is, or is the product of, a selection for reduced levels of GDP-fucose. In another embodiment, the manipulation is, or is the product of, a selection for reduced fucosylation of a glycoprotein. In another embodiment, the manipulation comprises contact with, or inclusion in or on the cell or batch of cultured cells, of an exogenous inhibitor of an enzyme involved in GDP-fucose biosynthesis, e.g., a specific or non-specific inhibitor. 
         [0492]    In an embodiments, an inhibitor, e.g., an inhibitor of GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose, is used, e.g., in the culture medium, to lower the levels of the GDP-fucose. In an embodiment the inhibitor can be guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, or p-chloromercuriphenylsulfonate EDTA. The inhibitor can be used with a cell which is mutant or wildtype for one or more GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0493]    In an embodiment the media contains a substance that can increase the level of GDP-fucose, e.g., butyrate or fucose. Such media can be used, e.g., with a cell having a mutation that eliminates or decreased the activity of one or more of GMD, FX, fucokinase, GFPP, GDP-synthetase, a fucosyltransferase or a GDP-Fucose transporter. 
         [0494]    In one aspect, the invention features a method of making a glycoprotein having reduced fucosylation, comprising:
       (a) providing, acknowledging, selecting, accepting, or memorializing a defined, desired or preselected glycan structure having reduced fucosylation for the glycoprotein,   (b) optionally providing a cell manipulated to decrease the level of fucosylation or fucose-GDP,   (c) culturing a cell manipulated to decrease the level of fucosylation or fucose-GDP, e.g., to form a batch of cultured cells, and   (d) isolating from the cell or batch of cultured cells a glycoprotein having the desired glycan structure,
 
thereby making a glycoprotein.
       
 
         [0499]    In one aspect, the invention features method of making a glycoprotein, comprising: 
         [0500]    providing, acknowledging, selecting, accepting, or memorializing a defined, desired or preselected glycan structure having reduced fucosylation for the glycoprotein, chosen, e.g., from Table 1; 
         [0501]    optionally, providing, acknowledging, selecting, accepting, or memorializing a manipulation described herein; 
         [0502]    culturing a cell having the manipulation, e.g., to form a batch of cultured cells;
       isolating from the cell or batch of cultured cells a glycoprotein having the desired glycan structure,
 
thereby making a glycoprotein.
       
 
         [0504]    In one aspect, the invention features method of formulating a pharmaceutical composition comprising: 
         [0505]    contacting a glycoprotein made by a method described herein with a pharmaceutically acceptable substance, e.g., an excipient or diluent. 
         [0506]    In one aspect, the invention features pharmaceutical preparation of a glycoprotein described herein or made by a method described herein, wherein the glycoprotein is selected from Table 1. 
         [0507]    Any step that generates information in a method described herein, e.g., a selection, analysis, comparison with a reference, or other evaluation or determination, can be memorialized, for example, by entry into a computer database. Such information can further be compared to a reference, or itself serve as a reference, for an evaluation made in the process. 
     
    
     DETAILED DESCRIPTION 
       [0508]    The drawings are first described. 
         [0509]      FIG. 1  is a plot of increasing amount of fucosylation on a glycoprotein produced by a cell (as a percentage of a cell without manipulation) (Y axis) against decreasing cellular GDP-fucose in the cell (as a percentage of a cell without manipulation). The plot shows a non-linear relationship indicative of a threshold relationship. E.g., reducing parental GDP-fucose levels by 20% gives little reduction in the amount of fucosylation. Reduction of more than 20% in GDP-fucose levels produced significant further reduction in glycosylation. Point A on the plot shows the point at which reduction in GDP-fucose begins to result in a significant reduction in fucosylation. Point B on the plot shows the point at which further reduction in GDP-fucose fails to result in further significant reduction in fucosylation. The region between points B and C is an optimal range. [&gt;20% and &lt;80% of parental GDP-fucose levels, e.g., &gt;40% and &lt;65% of parental GDP-fucose levels.] 
         [0510]      FIG. 2  is a depiction of glycan profiles from glycoproteins expressed from wild type CHO cells (top) and Lec 13.6 A cells (bottom). Data are negative mode MALDI spectra with the most abundant glycans indicated by structure. As indicated, glycans from the Lec 13.6 A cells have very low levels of fucosylation. 
       DEFINITIONS 
       [0511]    “Branched fucose” as used herein refers to a fucose moiety that is attached via an cd-3 or cd-4 linkage to an N-acetylglucosamine sugar of an N-linked or O-linked glycan component. 
         [0512]    “Core fucose” as used herein refers to a fucose moiety that is attached via an cd-6 linkage to the N-acetylglucosamine sugar that is directly attached to the asparagine amino acid in an N-linked glycan component. 
         [0513]    “Culturing” as used herein refers to placing a cell, e.g., a vertebrate, mammalian or rodent cell, under conditions that allow for at least some of the steps for the production of a glycoprotein to proceed. In embodiments, the conditions are sufficient to allow the glycosylation process to be completed. In embodiments, the conditions are sufficient to allow all of the steps, e.g., through secretion, to occur. Culturing refers to cultures of cells, cell lines, and populations of cells. The cells can be eukaryotic or a prokaryotic cells, e.g., animal, plant, yeast, fungal, insect or bacterial cells. In embodiments, culturing refers to in vitro culture of cells, e.g., primary or secondary cell lines. 
         [0514]    “Glycan complement” as used herein refers to all of the glycan components of a glycoprotein. In the case of a protein having a single glycosylation site, the glycan component attached thereto forms the glycan complement. In the case of a protein having more than one glycosylation site, the glycan complement is made up of the glycan components attached at all of the sites. The N-linked glycan complement refers to all of the N-linked glycan components of a protein. The O-linked glycan complement refers to all of the O-linked glycan components of a protein. A “component of the glycan complement” refers to a subset of the glycan components making up the glycan complement, e.g., one or more glycan components attached to its or their respective glycosylation site or sites. 
         [0515]    “Glycan component” as used herein refers to a sugar moiety, e.g., a monosaccharide, oligosaccharide or polysaccharide (e.g., a disaccharide, trisaccharide, tetrasaccharide, etc.) attached to a protein at one site. In embodiments the attachment is covalent and the glycan component is N- or O-linked to the protein. Glycan components can be chains of monosaccharides attached to one another via glycosidic linkages. Glycan components can be linear or branched. Fucose moieties are typically attached to an N-acetylglucosamine sugar of an N-linked or O-linked glycan component via an cd-3, cd-4 or cd-6 linkage. 
         [0516]    “Glycan structure” as used herein refers to the structure of a glycan complement, component of a glycan complement, or glycan component. In embodiments it refers to one or more of the placement and number of fucosyl moieties. 
         [0517]    A glycan structure can be described in terms of a comparison of the presence, absence or amount of a first glycan structure to a second glycan structure, for example, the presence, absence or amount of fucose relative to the presence, absence or amount of some other component. In other examples, the presence, absence or amount of fucose can be compared, e.g., to the presence, absence or amount of a sialic acid derivative such as N-glycolylneuraminic acid. 
         [0518]    Glycan structures can be described, identified or assayed in a number of ways. A glycan structure can be described, e.g., in defined structural terms, e.g., by chemical name, or by a functional or physical property, e.g., by molecular weight or by a parameter related to purification or separation, e.g., retention time of a peak in a column or other separation device. In embodiments a glycan structure can, by way of example, be a peak or other fraction (representing one or more species) from glycan structures derived from a glycoprotein, e.g., from an enzymatic digest. 
         [0519]    “Manipulation” as used herein can be any of a cell/activity-based manipulation, an envirocultural manipulation, or a selected functional manipulation. In general a manipulation is induced, selected, isolated, engineered, or is otherwise the product of the “hand of man.” 
         [0520]    A “cell/activity-based manipulation” as used herein refers to a property of a cell that decreases the level of GDP-fucose activity in a cell, e.g., which decreases the level of activity of an enzyme involved in GDP-fucose biosynthesis. Decreased means by comparison with a cell that is not subject to the cell/activity-based manipulation. 
         [0521]    Examples of cell/activity-based manipulations include: 
         [0522]    the presence in or on the cell of an exogenous inhibitor (e.g., an siRNA or a chemical inhibitor) of the activity of an enzyme involved in GDP-fucose biosynthesis; or 
         [0523]    a mutation or other genetic event that inhibits the activity of an enzyme involved in GDP-fucose biosynthesis. In some embodiments a cell/activity-based manipulation excludes genetic lesions, e.g., genetic knock-outs, discussed elsewhere herein. 
         [0524]    An “envirocultural manipulation” as used herein refers to a property of the culture conditions, e.g., of the culture medium, that lowers GDP-fucose level and results in a decrease in transfer of a fucose moiety to a glycoprotein. Examples include the modulation of salt or ion concentrations in the culture medium. Specific examples of media conditions that will lead to altered levels of GDP-fucose include but are not limited to altering the levels of cobalt, butyrate, fucose, guanosine, and manganese. 
         [0525]    A selected functional manipulation is a physical characteristic or property characterized, e.g., by the process that gave rise to it, e.g., a cell that was placed under selective conditions that result in the cell being able to produce a glycoprotein having a glycan structure characterized by a reduced GDP-fucose level, wherein the underlying basis for the ability to produce said glycoprotein having a glycan structure may or may not be known or characterized. 
         [0526]    “Reduced fucosylation” relates to the amount or frequency of fucosylation. With regard to a single molecule, it means fewer fucose moieties, e.g., as compared to a reference, e.g., a protein made by a cell without the manipulation that gave rise to reduced fucosylation. With regard to a plurality of molecules, e.g., a pharmaceutically acceptable preparation, it can mean fewer fucose moieties on the molecules of the plurality (e.g., as compared to a reference, e.g., the plurality made by cells without the manipulation that gave rise to reduced fucosylation). The comparison can be with regard to all fucosylation sites on the subject molecule or with regard to the fucosylation at one or more specific sites. Reduced fucosylation can mean reduced occupancy by, or presence of, a fucosyl moiety at a selected site, e.g., as compared to a reference preparation, e.g., a reference preparation made by cells without the manipulation that gave rise to reduced fucosylation. 
       Regulation of Glycosylation 
       [0527]    Glycosylation is a nonlinear non-template driven process. To this end, regulation of a particular glycan structure may be due to a number of orthogonal inputs such as precursor levels, donor levels, and transferase levels to name a few. Glycosylation of proteins can have dramatic effect on their activities, such as regulating receptor affinity, regulating bioavailability, or altering immunogenicity. For example, the presence of core fucosylation on an antibody may significantly attenuate antibody-dependent cell-mediated cytotoxicity (ADCC). 
         [0528]    Eukaryotic glycosylation occurs in the endoplasmic reticulum (ER) and Golgi through a stepwise process in which one monosaccharide is added through the activity of a glycosyltransferase, utilizing an activated sugar nucleotide as the donor molecule. The graphic below illustrates this with GDP-fucose. 
         [0000]    
       
                 
         
             
             
         
       
     
         [0529]    It should be noted that fucose can be added to a glycan structure at various points during the diversification process. This is one example of a glycan structure that may be fucosylated. 
       GDP-Fucose Biosynthesis 
       [0530]    Two pathways have been described for synthesis of GDP-fucose in the cytosol of essentially all mammalian cells, the de novo pathway and the salvage pathway. The de novo pathway transforms GDP-mannose to GDP-fucose via three enzymatic reactions carried out by two proteins, GDP-mannose 4,6-dehydratase (GMD) and GDP-keto-6-deoxymannose-3,5-epimerase-4-reductase (also known as the FX protein or tissue specific transplantation antigen P35B) (Scheme 1). The salvage pathway synthesizes GDP-fucose from free fucose derived from extracellular or lysosomal sources via the reactions of two proteins, a fucose kinase (fucokinase) followed by either GDP-fucose pyrophosphorylase (GFPP) (also known as fucose-1-phosphate guanylyltransferase) or GDP-fucose synthetase (Scheme 2). Quantitative studies of fucose metabolism in HeLa cells indicate that greater than 90% of GDP-fucose is derived from the de novo pathway (Yurchenco and Atkinson,  Biochemistry  14(14):3107-14, 1975; Yurchenco and Atkinson,  Biochemistry  16(5):944-53, 1977). 
         [0000]    
       
                 
         
             
             
         
       
     
         [0531]    Methods of regulating fucosylation by modulating levels of GDP-fucose, e.g., lowering GDP-fucose levels below a threshold level, are disclosed herein. In some embodiments this may involve the use of inhibitors of enzymes critical for GDP-fucose biosynthesis, such as GMD, FX, fucose kinase, GFPP and/or GDP-fucose synthetase. 
         [0532]    Exemplary proteins involved in GDP-fucose biosynthesis include the following: 
         [0000]    
       
         
               
             
           
               
                 Protein sequence of human GDP-mannose 4,6- 
               
               
                 dehydratase 
               
               
                 (SEQ ID NO: 1) 
               
               
                 MAHAPARCPSARGSGDGEMGKPRNVALITGITGQDGSYLAEFLLEKGYEV 
               
               
                   
               
               
                 HGIVRRSSSFNTGRIEHLYKNPQAHIEGNMKLHYGDLTDSTCLVKIINEV 
               
               
                   
               
               
                 KPTEIYNLGAQSHVKISFDLAEYTADVDGVGTLRLLDAVKTCGLINSVKF 
               
               
                   
               
               
                 YQASTSELYGKVQEIPQKETTPFYPRSPYGAAKLYAYWIVVNFREAYNLF 
               
               
                   
               
               
                 AVNGILFNHESPRRGANFVTRKISRSVAKIYLGQLECFSLGNLDAKRDWG 
               
               
                   
               
               
                 HAKDYVEAMWLMLQNDEPEDFVIATGEVHSVREFVEKSFLHIGKTIVWEG 
               
               
                   
               
               
                 KNENEVGRCKETGKVHVTVDLKYYRPTEVDFLQGDCTKAKQKLNWKPRVA 
               
               
                   
               
               
                 FDELVREMVHADVELMRTNPNA 
               
               
                   
               
               
                 GenBank Accession No. NP_001491 (GenBank version 
               
               
                   
               
               
                 dated 10-DEC-2008) 
               
               
                   
               
               
                 mRNA sequence of human GDP-mannose 4,6-dehydratase 
               
               
                 (SEQ ID NO: 2) 
               
               
                 ATGGCACACGCACCGGCACGCTGCCCCAGCGCCCGGGGCTCCGGGGACGG 
               
               
                   
               
               
                 CGAGATGGGCAAGCCCAGGAACGTGGCGCTCATCACCGGTATCACAGGCC 
               
               
                   
               
               
                 AGGATGGTTCCTACCTGGCTGAGTTCCTGCTGGAGAAAGGCTATGAGGTC 
               
               
                   
               
               
                 CATGGAATTGTACGGCGGTCCAGTTCATTTAATACGGGTCGAATTGAGCA 
               
               
                   
               
               
                 TCTGTATAAGAATCCCCAGGCTCACATTGAAGGAAACATGAAGTTGCACT 
               
               
                   
               
               
                 ATGGCGATCTCACTGACAGTACCTGCCTTGTGAAGATCATTAATGAAGTA 
               
               
                   
               
               
                 AAGCCCACAGAGATCTACAACCTTGGAGCCCAGAGCCACGTCAAAATTTC 
               
               
                   
               
               
                 CTTTGACCTCGCTGAGTACACTGCGGACGTTGACGGAGTTGGCACTCTAC 
               
               
                   
               
               
                 GACTTCTAGATGCAGTTAAGACTTGTGGCCTTATCAACTCTGTGAAGTTC 
               
               
                   
               
               
                 TACCAAGCCTCAACAAGTGAACTTTATGGGAAAGTGCAGGAAATACCCCA 
               
               
                   
               
               
                 GAAGGAGACCACCCCTTTCTATCCCCGGTCACCCTATGGGGCAGCAAAAC 
               
               
                   
               
               
                 TCTATGCCTATTGGATTGTGGTGAACTTCCGTGAGGCGTATAATCTCTTT 
               
               
                   
               
               
                 GCAGTGAACGGCATTCTCTTCAATCATGAGAGTCCCAGAAGAGGAGCTAA 
               
               
                   
               
               
                 TTTCGTTACTCGAAAAATTAGCCGGTCAGTAGCTAAGATTTACCTTGGAC 
               
               
                   
               
               
                 AACTGGAATGTTTCAGTTTGGGAAATCTGGATGCCAAACGAGATTGGGGC 
               
               
                   
               
               
                 CATGCCAAGGACTATGTGGAGGCTATGTGGTTGATGTTGCAGAATGATGA 
               
               
                   
               
               
                 GCCGGAGGACTTCGTTATAGCTACTGGGGAGGTCCATAGTGTCCGGGAAT 
               
               
                   
               
               
                 TTGTCGAGAAATCATTCTTGCACATTGGAAAAACCATTGTGTGGGAAGGA 
               
               
                   
               
               
                 AAGAATGAAAATGAAGTGGGCAGATGTAAAGAGACCGGCAAAGTTCACGT 
               
               
                   
               
               
                 GACTGTGGATCTCAAGTACTACCGGCCAACTGAAGTGGACTTTCTGCAGG 
               
               
                   
               
               
                 GCGACTGCACCAAAGCGAAACAGAAGCTGAACTGGAAGCCCCGGGTCGCT 
               
               
                   
               
               
                 TTCGATGAGCTGGTGAGGGAGATGGTGCACGCCGACGTGGAGCTCATGAG 
               
               
                   
               
               
                 GACAAACCCCAATGCCTGA 
               
               
                   
               
               
                 GenBank Accession No. NM_001500 (GenBank version 
               
               
                   
               
               
                 dated 10-DEC-2008) 
               
               
                   
               
               
                 Protein sequence of mouse GDP-mannose 4,6- 
               
               
                 dehydratase 
               
               
                 (SEQ ID NO: 3) 
               
               
                 MAQAPAKCPSYPGSGDGEMGKLRKVALITGITGQDGSYLAEFLLEKGYEV 
               
               
                   
               
               
                 HGIVRRSSSFNTGRIEHLYKNPQAHIEGNMKLHYGDLTDSTCLVKIINEV 
               
               
                   
               
               
                 KPTEIYNLGAQSHVKISFDLAEYTADVDGVGTLRLLDAIKTCGLINSVKF 
               
               
                   
               
               
                 YQASTSELYGKVQEIPQKETTPFYPRSPYGAAKLYAYWIVVNFREAYNLF 
               
               
                   
               
               
                 AVNGILFNHESPRRGANFVTRKISRSVAKIYLGQLECFSLGNLDAKRDWG 
               
               
                   
               
               
                 HAKDYVEAMWLMLQNDEPEDFVIATGEVHSVREFVEKSFMHIGKTIVWEG 
               
               
                   
               
               
                 KNENEVGRCKETGKVHVTVDLKYYRPTEVDFLQGDCSKAQQKLNWKPRVA 
               
               
                   
               
               
                 FDELVREMVQADVELMRTNPNA 
               
               
                   
               
               
                 GenBank Accession No. NP_666153 (GenBank version 
               
               
                   
               
               
                 dated 18-APR-2009) 
               
               
                   
               
               
                 mRNA sequence of mouse GDP-mannose 4,6-dehydratase 
               
               
                 (SEQ ID NO: 4) 
               
               
                 ATGGCTCAAGCTCCCGCTAAGTGCCCGAGCTACCCGGGCTCCGGGGATGG 
               
               
                   
               
               
                 CGAGATGGGCAAGCTCAGGAAGGTGGCTCTCATCACTGGCATCACCGGAC 
               
               
                   
               
               
                 AGGATGGTTCGTACTTGGCAGAATTCCTGTTGGAGAAAGGGTACGAGGTC 
               
               
                   
               
               
                 CATGGAATAGTACGGCGATCTAGTTCATTTAATACAGGTCGAATTGAACA 
               
               
                   
               
               
                 TTTATATAAGAATCCTCAGGCTCATATTGAAGGAAACATGAAGTTGCACT 
               
               
                   
               
               
                 ATGGTGACCTCACTGACAGCACCTGCCTAGTGAAAATCATCAATGAAGTC 
               
               
                   
               
               
                 AAGCCTACAGAGATCTATAATCTTGGAGCCCAGAGCCATGTCAAGATCTC 
               
               
                   
               
               
                 CTTTGACTTAGCTGAGTACACCGCAGATGTTGATGGCGTTGGCACCTTGC 
               
               
                   
               
               
                 GGCTTCTGGATGCAATTAAAACTTGTGGCCTTATAAATTCTGTGAAGTTC 
               
               
                   
               
               
                 TACCAGGCCTCAACAAGTGAACTTTATGGAAAAGTGCAGGAAATACCCCA 
               
               
                   
               
               
                 GAAGGAGACCACACCTTTCTATCCGAGGTCACCCTATGGAGCAGCCAAAC 
               
               
                   
               
               
                 TCTATGCCTATTGGATTGTGGTGAATTTCCGTGAAGCTTATAATCTCTTT 
               
               
                   
               
               
                 GCAGTGAATGGAATTCTCTTCAATCATGAGAGTCCCAGAAGAGGAGCTAA 
               
               
                   
               
               
                 TTTTGTTACTCGAAAAATTAGCCGGTCAGTAGCTAAGATTTACCTTGGAC 
               
               
                   
               
               
                 AACTGGAATGTTTCAGCTTGGGAAATCTGGATGCCAAACGAGACTGGGGC 
               
               
                   
               
               
                 CATGCCAAGGACTATGTAGAGGCTATGTGGCTCATGTTGCAGAATGATGA 
               
               
                   
               
               
                 GCCAGAGGACTTTGTCATAGCTACTGGGGAAGTTCACAGTGTCCGTGAAT 
               
               
                   
               
               
                 TTGTTGAAAAGTCATTCATGCACATCGGAAAAACCATTGTGTGGGAAGGA 
               
               
                   
               
               
                 AAGAATGAAAATGAAGTGGGCAGATGTAAAGAGACCGGCAAAGTTCACGT 
               
               
                   
               
               
                 GACTGTGGATCTGAAATACTACCGACCGACTGAAGTGGACTTTCTGCAGG 
               
               
                   
               
               
                 GAGACTGCTCCAAGGCTCAGCAGAAGCTAAACTGGAAGCCCCGCGTTGCC 
               
               
                   
               
               
                 TTTGACGAGCTGGTGAGGGAGATGGTGCAGGCCGACGTGGAGCTCATGAG 
               
               
                   
               
               
                 GACCAACCCCAACGCTTGA 
               
               
                   
               
               
                 GenBank Accession No. NM_146041 (GenBank version 
               
               
                   
               
               
                 dated 18-APR-2009) 
               
               
                   
               
               
                 Protein sequence of rat GDP-mannose 4,6- 
               
               
                 dehydratase 
               
               
                 (SEQ ID NO: 5) 
               
               
                 MAHAPASCRRYPGSGDGEMGKLRKVALITGITGQDGSYLAEFLLEKGYEV 
               
               
                   
               
               
                 HGIVRRSSSFNTGRIEHLYKNPQAHIEGNMKLHYGDLTDSTCLVKIINEV 
               
               
                   
               
               
                 KPTEIYNLGAQSHVKISFDLAEYTADVDGVGTLRLLDAIKTCGLINSVKF 
               
               
                   
               
               
                 YQASTSELYGKVQEIPQKETTPFYPRSPYGAAKLYAYWIVVNFREAYNLF 
               
               
                   
               
               
                 AVNGILFNHESPRRGANFVTRKISRSVAKIYLGQLECFSLGNLDAKRDWG 
               
               
                   
               
               
                 HAKDYVEAMWLMLQNDEPEDFVIATGEVHSVREFVEKSFMHIGKTIVWEG 
               
               
                   
               
               
                 KNENEVGRCKETGKIHVTVDLKYYRPTEVDFLQGDCSKAQQKLNWKPRVA 
               
               
                   
               
               
                 FDELVREMVQADVELMRTNPNA 
               
               
                   
               
               
                 GenBank Accession No. NP_001034695 (GenBank 
               
               
                   
               
               
                 version dated 18-APR-2009) 
               
               
                   
               
               
                 mRNA sequence of rat GDP-mannose 4,6-dehydratase 
               
               
                 (SEQ ID NO: 6) 
               
               
                 ATGGCCCACGCTCCCGCTAGCTGCCGGAGATACCCGGGCTCCGGGGATGG 
               
               
                   
               
               
                 CGAGATGGGCAAGCTCAGGAAGGTAGCTCTCATCACCGGCATCACTGGCC 
               
               
                   
               
               
                 AGGATGGTTCATACTTGGCAGAATTCCTGCTGGAGAAAGGATACGAGGTC 
               
               
                   
               
               
                 CATGGAATAGTACGGCGATCTAGTTCATTTAATACAGGTCGAATTGAACA 
               
               
                   
               
               
                 TTTATATAAGAATCCTCAGGCTCATATTGAAGGAAACATGAAGTTGCACT 
               
               
                   
               
               
                 ATGGCGACCTGACTGACAGCACCTGCCTGGTGAAAATCATCAATGAAGTG 
               
               
                   
               
               
                 AAGCCTACAGAGATCTACAATCTTGGCGCTCAGAGCCATGTCAAGATCTC 
               
               
                   
               
               
                 CTTTGACTTAGCTGAATACACCGCAGACGTTGATGGAGTTGGCACCTTGC 
               
               
                   
               
               
                 GGCTTCTGGATGCAATTAAAACTTGCGGCCTTATAAATTCTGTGAAGTTC 
               
               
                   
               
               
                 TACCAGGCCTCGACAAGTGAACTTTATGGAAAAGTTCAGGAAATACCCCA 
               
               
                   
               
               
                 GAAAGAGACCACACCTTTCTATCCGAGGTCACCCTATGGAGCCGCCAAGC 
               
               
                   
               
               
                 TCTATGCCTATTGGATTGTGGTGAATTTCCGTGAAGCTTATAATCTCTTT 
               
               
                   
               
               
                 GCAGTGAATGGCATTCTCTTCAATCACGAGAGCCCCAGAAGAGGAGCTAA 
               
               
                   
               
               
                 TTTTGTTACTCGAAAAATTAGCCGGTCAGTAGCTAAGATTTACCTTGGAC 
               
               
                   
               
               
                 AACTGGAATGTTTCAGTTTGGGAAATCTGGATGCCAAACGAGACTGGGGC 
               
               
                   
               
               
                 CATGCCAAGGACTATGTAGAGGCTATGTGGCTGATGTTGCAAAATGATGA 
               
               
                   
               
               
                 GCCGGAGGACTTTGTCATAGCTACTGGGGAAGTTCACAGTGTCCGTGAAT 
               
               
                   
               
               
                 TTGTTGAAAAATCATTCATGCACATTGGAAAAACCATTGTGTGGGAAGGA 
               
               
                   
               
               
                 AAGAATGAAAATGAAGTAGGCAGATGTAAGGAGACCGGCAAAATTCACGT 
               
               
                   
               
               
                 GACTGTGGATCTGAAATACTACCGACCGACTGAAGTGGACTTTCTACAGG 
               
               
                   
               
               
                 GAGACTGCTCCAAGGCTCAGCAGAAACTGAACTGGAAACCCCGCGTTGCC 
               
               
                   
               
               
                 TTCGATGAGCTGGTGAGAGAGATGGTGCAGGCCGACGTGGAGCTCATGAG 
               
               
                   
               
               
                 GACCAACCCCAACGCTTGA 
               
               
                   
               
               
                 GenBank Accession No. NM_001039606 (GenBank 
               
               
                   
               
               
                 version dated 18-APR-2009) 
               
               
                   
               
               
                 Protein sequence of Chinese hamster GDP-mannose 
               
               
                 4,6-dehydratase 
               
               
                 (SEQ ID NO: 7) 
               
               
                 MAHAPARCPSARGSGDGEMGKPRNVALITGITGQDGSYLAEFLLEKGYEV 
               
               
                   
               
               
                 HGIVRRSSSFNTGRIEHLYKNPQAHIEGNMKLHYGDLTDSTCLVKIINEV 
               
               
                   
               
               
                 KPTEIYNLGAQSHVKISFDLAEYTADVDGVGTLRLLDAVKTCGLINSVKF 
               
               
                   
               
               
                 YQASTSELYGKVQEIPQKETTPFYPRSPYGAAKLYAYWIVVNFREAYNLF 
               
               
                   
               
               
                 AVNGILFNHESPRRGANFVTRKISRSVAKIYLGQLECFSLGNLDAKRDWG 
               
               
                   
               
               
                 HAKDYVEAMWLMLQNDEPEDFVIATGEVHSVREFVEKSFLHIGKTIVWEG 
               
               
                   
               
               
                 KNENEVGRCKETGKVHVTVDLKYYRPTEVDFLQGDCTKAKQKLNWKPRVA 
               
               
                   
               
               
                 FDELVREMVHADVELMRTNPNA 
               
               
                   
               
               
                 GenBank Accession No. Q8K3X3 (GenBank version 
               
               
                   
               
               
                 dated 20-JAN-2009) 
               
               
                   
               
               
                 mRNA sequence of Chinese hamster GDP-mannose 
               
               
                 4,6-dehydratase 
               
               
                 (SEQ ID NO: 8) 
               
               
                 agactgtggcggccgctgcagctccgtgaggcgactggcgcgcgcaccca 
               
               
                   
               
               
                 cgtctctgtcggcccgctgccggttccacggttccactcctccttccact 
               
               
                   
               
               
                 cggctgcacgctcacccgcccgcggcgacATGGCTCACGCTCCCGCTAGC 
               
               
                   
               
               
                 TGCCCGAGCTCCAGGAACTCTGGGGACGGCGATAAGGGCAAGCCCAGGAA 
               
               
                   
               
               
                 GGTGGCGCTCATCACGGGCATCACCGGCCAGGATGGCTCATACTTGGCAG 
               
               
                   
               
               
                 AATTCCTGCTGGAGAAAGGATACGAGGTTCATGGAATTGTACGGCGATCC 
               
               
                   
               
               
                 AGTTCATTTAATACAGGTCGAATTGAACATTTATATAAGAATCCACAGGC 
               
               
                   
               
               
                 TCATATTGAAGGAAACATGAAGTTGCACTATGGTGACCTCACCGACAGCA 
               
               
                   
               
               
                 CCTGCCTAGTAAAAATCATCAATGAAGTCAAACCTACAGAGATCTACAAT 
               
               
                   
               
               
                 CTTGGTGCCCAGAGCCATGTCAAGATTTCCTTTGACTTAGCAGAGTACAC 
               
               
                   
               
               
                 TGCAGATGTTGATGGAGTTGGCACCTTGCGGCTTCTGGATGCAATTAAGA 
               
               
                   
               
               
                 CTTGTGGCCTTATAAATTCTGTGAAGTTCTACCAGGCCTCAACTAGTGAA 
               
               
                   
               
               
                 CTGTATGGAAAAGTGCAAGAAATACCCCAGAAAGAGACCACCCCTTTCTA 
               
               
                   
               
               
                 TCCAAGGTCGCCCTATGGAGCAGCCAAACTTTATGCCTATTGGATTGTAG 
               
               
                   
               
               
                 TGAACTTTCGAGAGGCTTATAATCTCTTTGCGGTGAACGGCATTCTCTTC 
               
               
                   
               
               
                 AATCATGAGAGTCCTAGAAGAGGAGCTAATTTTGTTACTCGAAAAATTAG 
               
               
                   
               
               
                 CCGGTCAGTAGCTAAGATTTACCTTGGACAACTGGAATGTTTCAGTTTGG 
               
               
                   
               
               
                 GAAATCTGGACGCCAAACGAGACTGGGGCCATGCCAAGGACTATGTCGAG 
               
               
                   
               
               
                 GCTATGTGGCTGATGTTACAAAATGATGAACCAGAGGACTTTGTCATAGC 
               
               
                   
               
               
                 TACTGGGGAAGTTCATAGTGTCCGTGAATTTGTTGAGAAATCATTCATGC 
               
               
                   
               
               
                 ACATTGGAAAGACCATTGTGTGGGAAGGAAAGAATGAAAATGAAGTGGGC 
               
               
                   
               
               
                 AGATGTAAAGAGACCGGCAAAATTCATGTGACTGTGGATCTGAAATACTA 
               
               
                   
               
               
                 CCGACCAACTGAAGTGGACTTCCTGCAGGGAGACTGCTCCAAGGCGCAGC 
               
               
                   
               
               
                 AGAAACTGAACTGGAAGCCCCGCGTTGCCTTTGACGAGCTGGTGAGGGAG 
               
               
                   
               
               
                 ATGGTGCAAGCCGATGTGGAGCTCATGAGAACCAACCCCAACGCCTGAgc 
               
               
                   
               
               
                 acctctacaaaaaattcgcgagacatggactatggtgcagagccagccaa 
               
               
                   
               
               
                 ccagagtccagccactcctgagaccatcgaccataaaccctcgactgcct 
               
               
                   
               
               
                 gtgtcgtccccacagctaagagctgggccacaggtttgtgggcaccagga 
               
               
                   
               
               
                 cggggacactccagagctaaggccacttcgcttttgtcaaaggctcctct 
               
               
                   
               
               
                 gaatgattttgggaaatcaagaagtttaaaatcacatactcattttactt 
               
               
                   
               
               
                 gaaattatgtcactagacaacttaaatttttgagtcttgagattgttttt 
               
               
                   
               
               
                 ctcttttcttattaaatgatctttctatgaaccagcaaaaaaaaaaaaaa 
               
               
                   
               
               
                 aaaaaa 
               
               
                   
               
               
                 GenBank Accession No. AF525364 (GenBank version 
               
               
                   
               
               
                 dated 04-AUG-2002) 
               
               
                   
               
               
                 Protein sequence of human GDP-keto-6-deoxymannose 
               
               
                 3,5-epimerase, 4-reductase (FX protein, tissue 
               
               
                 specific transplantation antigen P35B) 
               
               
                 (SEQ ID NO: 9) 
               
               
                 MGEPQGSMRILVTGGSGLVGKAIQKVVADGAGLPGEDWVFVSSKDADLTD 
               
               
                   
               
               
                 TAQTRALFEKVQPTHVIHLAAMVGGLFRNIKYNLDFWRKNVHMNDNVLHS 
               
               
                   
               
               
                 AFEVGARKVVSCLSTCIFPDKTTYPIDETMIHNGPPHNSNFGYSYAKRMI 
               
               
                   
               
               
                 DVQNRAYFQQYGCTFTAVIPTNVFGPHDNFNIEDGHVLPGLIHKVHLAKS 
               
               
                   
               
               
                 SGSALTVWGTGNPRRQFIYSLDLAQLFIWVLREYNEVEPIILSVGEEDEV 
               
               
                   
               
               
                 SIKEAAEAVVEAMDFHGEVTFDTTKSDGQFKKTASNSKLRTYLPDFRFTP 
               
               
                   
               
               
                 FKQAVKETCAWFTDNYEQARK 
               
               
                   
               
               
                 GenBank Accession No. NP_003304 (GenBank version 
               
               
                   
               
               
                 dated 10-DEC-2008) 
               
               
                   
               
               
                 mRNA sequence of human GDP-keto-6-deoxymannose 
               
               
                 3,5-epimerase, 4-reductase (FX protein, tissue 
               
               
                 specific transplantation antigen P35B) 
               
               
                 (SEQ ID NO: 10) 
               
               
                 ATGGGTGAACCCCAGGGATCCATGCGGATTCTAGTGACAGGGGGCTCTGG 
               
               
                   
               
               
                 GCTGGTAGGCAAAGCCATCCAGAAGGTGGTAGCAGATGGAGCTGGACTTC 
               
               
                   
               
               
                 CTGGAGAGGACTGGGTGTTTGTCTCCTCTAAAGACGCCGATCTCACGGAT 
               
               
                   
               
               
                 ACAGCACAGACCCGCGCCCTGTTTGAGAAGGTCCAACCCACACACGTCAT 
               
               
                   
               
               
                 CCATCTTGCTGCAATGGTGGGGGGCCTGTTCCGGAATATCAAATACAATT 
               
               
                   
               
               
                 TGGACTTCTGGAGGAAAAACGTGCACATGAACGACAACGTCCTGCACTCG 
               
               
                   
               
               
                 GCCTTTGAGGTGGGCGCCCGCAAGGTGGTGTCCTGCCTGTCCACCTGTAT 
               
               
                   
               
               
                 CTTCCCTGACAAGACGACCTACCCGATAGATGAGACCATGATCCACAATG 
               
               
                   
               
               
                 GGCCTCCCCACAACAGCAATTTTGGGTACTCGTATGCCAAGAGGATGATC 
               
               
                   
               
               
                 GACGTGCAGAACAGGGCCTACTTCCAGCAGTACGGCTGCACCTTCACCGC 
               
               
                   
               
               
                 TGTCATCCCCACCAACGTCTTCGGGCCCCACGACAACTTCAACATCGAGG 
               
               
                   
               
               
                 ATGGCCACGTGCTGCCTGGCCTCATCCACAAGGTGCACCTGGCCAAGAGC 
               
               
                   
               
               
                 AGCGGCTCGGCCCTGACGGTGTGGGGTACAGGGAATCCGCGGAGGCAGTT 
               
               
                   
               
               
                 CATATACTCGCTGGACCTGGCCCAGCTCTTTATCTGGGTCCTGCGGGAGT 
               
               
                   
               
               
                 ACAATGAAGTGGAGCCCATCATCCTCTCCGTGGGCGAGGAAGATGAGGTC 
               
               
                   
               
               
                 TCCATCAAGGAGGCAGCCGAGGCGGTGGTGGAGGCCATGGACTTCCATGG 
               
               
                   
               
               
                 GGAAGTCACCTTTGATACAACCAAGTCGGATGGGCAGTTTAAGAAGACAG 
               
               
                   
               
               
                 CCAGTAACAGCAAGCTGAGGACCTACCTGCCCGACTTCCGGTTCACACCC 
               
               
                   
               
               
                 TTCAAGCAGGCGGTGAAGGAGACCTGTGCTTGGTTCACTGACAACTACGA 
               
               
                   
               
               
                 GCAGGCCCGGAAGTGA 
               
               
                   
               
               
                 GenBank Accession No. NM_003313 (GenBank version 
               
               
                   
               
               
                 dated 10-DEC-2008) 
               
               
                   
               
               
                 Protein sequence of mouse GDP-keto-6-deoxymannose 
               
               
                 3,5-epimerase, 4-reductase (FX protein, tissue 
               
               
                 specific transplantation antigen P35B) 
               
               
                 (SEQ ID NO: 11) 
               
               
                 MGEPHGSMRILVTGGSGLVGRAIQKVVADGAGLPGEEWVFVSSKDADLTD 
               
               
                   
               
               
                 AAQTQALFQKVQPTHVIHLAAMVGGLFRNIKYNLDFWRKNVHINDNVLHS 
               
               
                   
               
               
                 AFEVGARKVVSCLSTCIFPDKTTYPIDETMIHNGPPHSSNFGYSYAKRMI 
               
               
                   
               
               
                 DVQNRAYFQQHGCTFTAVIPTNVFGPYDNFNIEDGHVLPGLIHKVHLAKS 
               
               
                   
               
               
                 SDSALTVWGTGKPRRQFIYSLDLARLFIWVLREYSEVEPIILSVGEEDEV 
               
               
                   
               
               
                 SIKEAAEAVVEAMDFNGEVTFDSTKSDGQYKKTASNGKLRSYLPDFRFTP 
               
               
                   
               
               
                 FKQAVKETCTWFTDNYEQARK 
               
               
                   
               
               
                 GenBank Accession No. NP_112478 (GenBank version 
               
               
                   
               
               
                 dated 10-MAY-2009) 
               
               
                   
               
               
                 mRNA sequence of mouse GDP-keto-6-deoxymannose 
               
               
                 3,5-epimerase, 4-reductase (FX protein, tissue 
               
               
                 specific transplantation antigen P35B) 
               
               
                 (SEQ ID NO: 12) 
               
               
                 ATGGGCGAACCCCATGGATCCATGAGGATCCTAGTGACAGGGGGCTCTGG 
               
               
                   
               
               
                 ACTGGTGGGTAGAGCCATCCAGAAGGTGGTTGCAGATGGGGCCGGCTTAC 
               
               
                   
               
               
                 CTGGAGAGGAATGGGTGTTTGTCTCCTCCAAAGATGCAGATCTGACGGAT 
               
               
                   
               
               
                 GCAGCCCAAACCCAAGCACTCTTCCAGAAAGTACAGCCCACCCACGTCAT 
               
               
                   
               
               
                 CCATCTCGCTGCAATGGTAGGCGGCCTTTTCCGGAATATCAAATACAACT 
               
               
                   
               
               
                 TGGATTTCTGGCGGAAAAACGTGCACATCAATGACAACGTCCTGCATTCG 
               
               
                   
               
               
                 GCCTTCGAGGTGGGCGCTCGCAAGGTGGTCTCCTGCCTGTCCACCTGCAT 
               
               
                   
               
               
                 CTTCCCTGACAAGACCACCTATCCTATTGACGAGACAATGATCCACAACG 
               
               
                   
               
               
                 GGCCGCCTCACAGCAGCAATTTCGGGTACTCATACGCCAAGAGGATGATT 
               
               
                   
               
               
                 GACGTGCAGAACAGAGCCTACTTCCAGCAGCACGGCTGTACCTTCACCGC 
               
               
                   
               
               
                 CGTCATCCCTACCAATGTCTTTGGGCCTTATGACAACTTCAACATCGAAG 
               
               
                   
               
               
                 ATGGCCACGTGCTACCCGGCCTCATCCATAAGGTGCACCTGGCCAAGAGT 
               
               
                   
               
               
                 AGTGACTCGGCCCTGACGGTGTGGGGTACAGGGAAGCCGCGGAGGCAGTT 
               
               
                   
               
               
                 CATCTACTCACTGGACCTCGCCCGGCTCTTCATCTGGGTCCTACGGGAGT 
               
               
                   
               
               
                 ACAGTGAGGTGGAGCCCATCATCCTCTCAGTGGGTGAGGAAGATGAAGTG 
               
               
                   
               
               
                 TCCATCAAGGAGGCAGCTGAGGCTGTAGTGGAGGCCATGGACTTCAATGG 
               
               
                   
               
               
                 GGAAGTCACTTTTGATTCAACAAAGTCAGATGGGCAATATAAGAAGACAG 
               
               
                   
               
               
                 CCAGCAATGGCAAGTTGCGGTCCTACTTGCCCGACTTCCGTTTCACACCC 
               
               
                   
               
               
                 TTCAAGCAGGCTGTGAAGGAAACCTGCACTTGGTTCACCGACAACTATGA 
               
               
                   
               
               
                 GCAGGCCCGGAAGTAA 
               
               
                   
               
               
                 GenBank Accession No. NM_031201 (GenBank version 
               
               
                   
               
               
                 dated 10-MAY-2009) 
               
               
                   
               
               
                 Protein sequence of rat GDP-keto-6-deoxymannose 
               
               
                 3,5-epimerase, 4-reductase (FX protein, tissue 
               
               
                 specific transplantation antigen P35B) 
               
               
                 (SEQ ID NO: 13) 
               
               
                 MGEPHGSMRILVTGGSGLVGRAIQKVVADGAGLPGEEWVFVSSKDADLTD 
               
               
                   
               
               
                 AAQTQALFQKVQPTHVIHLAAMVGGLFRNIKYNLDFWRKNVHINDNVLHS 
               
               
                   
               
               
                 AFEVGTRKVVSCLSTCIFPDKTTYPIDETMIHNGPPHSSNFGYSYAKRMI 
               
               
                   
               
               
                 DVQNRAYFQQHGCTFTSVIPTNVFGPYDNFNIEDGHVLPGLIHKVHLAKS 
               
               
                   
               
               
                 SGSALTVWGTGKPRRQFIYSLDLARLFIWVLREYNEVEPIILSVGEEDEV 
               
               
                   
               
               
                 SIKEAAEAVVEAMDFSGEVTFDSTKSDGQYKKTASNGKLRSYLPDFCFTP 
               
               
                   
               
               
                 FKQAVKETCAWFTENYEQARK 
               
               
                   
               
               
                 GenBank Accession No. NP_001120927 (GenBank 
               
               
                   
               
               
                 version dated 24-AUG-2008) 
               
               
                   
               
               
                 mRNA sequence of rat GDP-keto-6-deoxymannose 3,5- 
               
               
                 epimerase, 4-reductase (FX protein, tissue 
               
               
                 specific transplantation antigen P35B) 
               
               
                 (SEQ ID NO: 14) 
               
               
                 ATGGGTGAACCCCACGGATCCATGAGGATCCTAGTAACAGGGGGCTCTGG 
               
               
                   
               
               
                 ACTGGTGGGCAGAGCCATCCAGAAGGTGGTCGCAGATGGGGCCGGCTTGC 
               
               
                   
               
               
                 CTGGAGAGGAATGGGTGTTTGTCTCCTCCAAAGATGCAGATCTGACGGAT 
               
               
                   
               
               
                 GCAGCGCAAACCCAAGCTCTGTTCCAGAAGGTACAGCCCACCCACGTCAT 
               
               
                   
               
               
                 CCATCTTGCTGCAATGGTAGGCGGCCTTTTCCGGAATATTAAATACAACT 
               
               
                   
               
               
                 TGGATTTCTGGAGGAAGAACGTGCACATCAATGACAACGTCCTACATTCA 
               
               
                   
               
               
                 GCCTTCGAGGTGGGCACACGCAAGGTGGTCTCCTGCCTGTCCACCTGCAT 
               
               
                   
               
               
                 CTTCCCTGACAAGACCACCTATCCTATTGATGAGACCATGATCCACAACG 
               
               
                   
               
               
                 GGCCGCCTCACAGCAGCAATTTTGGGTACTCATATGCCAAGAGGATGATT 
               
               
                   
               
               
                 GACGTGCAGAACAGGGCCTACTTCCAGCAGCATGGCTGTACCTTCACCTC 
               
               
                   
               
               
                 TGTCATCCCTACCAATGTCTTTGGGCCTTACGACAACTTCAACATCGAAG 
               
               
                   
               
               
                 ATGGCCACGTGCTGCCGGGCCTCATCCATAAGGTGCACCTGGCCAAGAGC 
               
               
                   
               
               
                 AGTGGTTCAGCCTTGACTGTGTGGGGTACGGGGAAGCCGCGGAGACAGTT 
               
               
                   
               
               
                 CATCTACTCACTGGACCTAGCCCGGCTCTTCATCTGGGTCCTTCGGGAGT 
               
               
                   
               
               
                 ACAATGAGGTGGAGCCCATCATCCTCTCAGTGGGCGAGGAAGATGAAGTG 
               
               
                   
               
               
                 TCTATCAAGGAGGCAGCTGAGGCTGTGGTGGAGGCCATGGACTTCTCTGG 
               
               
                   
               
               
                 GGAAGTCACTTTTGATTCAACAAAGTCAGATGGGCAGTATAAGAAGACAG 
               
               
                   
               
               
                 CCAGCAATGGCAAGTTGCGGTCCTACTTGCCTGACTTCTGTTTCACACCC 
               
               
                   
               
               
                 TTCAAGCAGGCTGTGAAGGAAACTTGTGCTTGGTTCACTGAAAACTACGA 
               
               
                   
               
               
                 GCAGGCCCGGAAGTAA 
               
               
                   
               
               
                 GenBank Accession No. NM_001127455 (GenBank 
               
               
                   
               
               
                 version dated 24-AUG-2008) 
               
               
                   
               
               
                 Protein sequence of Chinese hamster GDP-keto-6- 
               
               
                 deoxymannose 3,5-epimerase, 4-reductase 
               
               
                 (SEQ ID NO: 15) 
               
               
                 MGEPQGSRRILVTGGSGLVGRAIQKVVADGAGLPGEEWVFVSSKDADLTD 
               
               
                   
               
               
                 AAQTQALFQKVQPTHVIHLAAMVGGLFRNIKYNLDFWRKNVHINDNVLHS 
               
               
                   
               
               
                 AFEVGTRKVVSCLSTCIFPDKTTYPIDETMIHNGPPHSSNFGYSYAKRMI 
               
               
                   
               
               
                 DVQNRAYFQQHGCTFTAVIPTNVFGPHDNFNIEDGHVLPGLIHKVHLAKS 
               
               
                   
               
               
                 NGSALTVWGTGKPRRQFIYSLDLARLFIWVLREYNEVEPIILSVGEEDEV 
               
               
                   
               
               
                 SIKEAAEAVVEAMDFCGEVTFDSTKSDGQYKKTASNGKLRAYLPDFRFTP 
               
               
                   
               
               
                 FKQAVKETCAWFTDNYEQARK 
               
               
                   
               
               
                 GenBank Accession No. Q8K3X2 (GenBank version 
               
               
                   
               
               
                 dated 20-JAN-2009) 
               
               
                   
               
               
                 mRNA sequence of Chinese hamster GDP-keto-6- 
               
               
                 deoxymannose 3,5-epimerase, 4-reductase (FX 
               
               
                 protein) 
               
               
                 (SEQ ID NO: 16) 
               
               
                 ccggaagtagctcttggactggtggaaccctgcgcaggtgcagcaacaAT 
               
               
                   
               
               
                 GGGTGAGCCCCAGGGATCCAGGAGGATCCTAGTGACAGGGGGCTCTGGAC 
               
               
                   
               
               
                 TGGTGGGCAGAGCTATCCAGAAGGTGGTCGCAGATGGCGCTGGCTTACCC 
               
               
                   
               
               
                 GGAGAGGAATGGGTGTTTGTCTCCTCCAAAGATGCAGATCTGACGGATGC 
               
               
                   
               
               
                 AGCACAAACCCAAGCCCTGTTCCAGAAGGTACAGCCCACCCATGTCATCC 
               
               
                   
               
               
                 ATCTTGCTGCAATGGTAGGAGGCCTTTTCCGGAATATCAAATACAACTTG 
               
               
                   
               
               
                 GATTTCTGGAGGAAGAATGTGCACATCAATGACAACGTCCTGCACTCAGC 
               
               
                   
               
               
                 TTTCGAGGTGGGCACTCGCAAGGTGGTCTCCTGCCTGTCCACCTGTATCT 
               
               
                   
               
               
                 TCCCTGACAAGACCACCTATCCTATTGATGAAACAATGATCCACAATGGT 
               
               
                   
               
               
                 CCACCCCACAGCAGCAATTTTGGGTACTCGTATGCCAAGAGGATGATTGA 
               
               
                   
               
               
                 CGTGCAGAACAGGGCCTACTTCCAGCAGCATGGCTGCACCTTCACTGCTG 
               
               
                   
               
               
                 TCATCCCTACCAATGTCTTTGGACCTCATGACAACTTCAACATTGAAGAT 
               
               
                   
               
               
                 GGCCATGTGCTGCCTGGCCTCATCCATAAGGTGCATCTGGCCAAGAGTAA 
               
               
                   
               
               
                 TGGTTCAGCCTTGACTGTTTGGGGTACAGGGAAACCACGGAGGCAGTTCA 
               
               
                   
               
               
                 TCTACTCACTGGACCTAGCCCGGCTCTTCATCTGGGTCCTGCGGGAGTAC 
               
               
                   
               
               
                 AATGAAGTTGAGCCCATCATCCTCTCAGTGGGCGAGGAAGATGAAGTCTC 
               
               
                   
               
               
                 CATTAAGGAGGCAGCTGAGGCTGTAGTGGAGGCCATGGACTTCTGTGGGG 
               
               
                   
               
               
                 AAGTCACTTTTGATTCAACAAAGTCAGATGGGCAGTATAAGAAGACAGCC 
               
               
                   
               
               
                 AGCAATGGCAAGCTTCGGGCCTACTTGCCTGATTTCCGTTTCACACCCTT 
               
               
                   
               
               
                 CAAGCAGGCTGTGAAGGAGACCTGTGCCTGGTTCACCGACAACTATGAGC 
               
               
                   
               
               
                 AGGCCCGGAAGTGAagcatgggacaagcgggtgctcagctggcaatgccc 
               
               
                   
               
               
                 agtcagtaggctgcagtctcatcatttgcttgtcaagaactgaggacagt 
               
               
                   
               
               
                 atccagcaacctgagccacatgctggtctctctgccagggggcttcatgc 
               
               
                   
               
               
                 agccatccagtagggcccatgtttgtccatcctcgggggaaggccagacc 
               
               
                   
               
               
                 aacaccttgtttgtctgcttctgccccaacctcagtgcatccatgctggt 
               
               
                   
               
               
                 cctgctgtcccttgtctagaaaccaataaaatggattttcataaaaaaaa 
               
               
                   
               
               
                 aaaaaaaaaaa 
               
               
                   
               
               
                 GenBank Accession No. AF525365 (GenBank version 
               
               
                   
               
               
                 dated 04-AUG-2002) 
               
               
                   
               
               
                 Protein sequence of human GDP fucose pyrophos- 
               
               
                 phorylase 
               
               
                 (SEQ ID NO: 17) 
               
               
                 MAAARDPPEVSLREATQRKLRRFSELRGKLVARGEFWDIVAITAADEKQE 
               
               
                   
               
               
                 LAYNQQLSEKLKRKELPLGVQYHVFVDPAGAKIGNGGSTLCALQCLEKLY 
               
               
                   
               
               
                 GDKWNSFTILLIHSGGYSQRLPNASALGKIFTALPLGNPIYQMLELKLAM 
               
               
                   
               
               
                 YIDFPLNMNPGILVTCADDIELYSIGEFEFIRFDKPGFTALAHPSSLTIG 
               
               
                   
               
               
                 TTHGVFVLDPFDDLKHRDLEYRSCHRFLHKPSIEKMYQFNAVCRPGNFCQ 
               
               
                   
               
               
                 QDFAGGDIADLKLDSDYVYTDSLFYMDHKSAKMLLAFYEKIGTLSCEIDA 
               
               
                   
               
               
                 YGDFLQALGPGATVEYTRNTSHVIKEESELVEMRQRIFHLLKGTSLNVVV 
               
               
                   
               
               
                 LNNSKFYHIGTTEEYLFYFTSDNSLKSELGLQSITFSIFPDIPECSGKTS 
               
               
                   
               
               
                 CIIQSILDSRCSVAPGSVVEYSRLGPDVSVGENCIISGSYILTKAALPAH 
               
               
                   
               
               
                 SFVCSLSLKMNRCLKYATMAFGVQDNLKKSVKTLSDIKLLQFFGVCFLSC 
               
               
                   
               
               
                 LDVWNLKVTEELFSGNKTCLSLWTARIFPVCSSLSDSVITSLKMLNAVKN 
               
               
                   
               
               
                 KSAFSLNSYKLLSIEEMLIYKDVEDMITYREQIFLEISLKSSLM 
               
               
                   
               
               
                 GenBank Accession No. AAC73005 (GenBank version 
               
               
                   
               
               
                 dated 12-NOV-1998) 
               
               
                   
               
               
                 mRNA sequence of human GDP fucose pyrophos- 
               
               
                 phorylase 
               
               
                 (SEQ ID NO: 18) 
               
               
                 ATGGCAGCTGCTAGGGACCCTCCGGAAGTATCGCTGCGAGAAGCCACCCA 
               
               
                   
               
               
                 GCGAAAATTGCGGAGGTTTTCCGAGCTAAGAGGCAAACTTGTAGCACGTG 
               
               
                   
               
               
                 GAGAATTCTGGGACATAGTTGCAATAACAGCGGCTGATGAAAAACAGGA 
               
               
                   
               
               
                 ACTTGCTTACAACCAACAGCTGTCAGAAAAGCTGAAAAGAAAGGAGTTAC 
               
               
                   
               
               
                 CCCTTGGAGTTCAATATCACGTTTTTGTGGATCCTGCTGGAGCCAAAATT 
               
               
                   
               
               
                 GGAAATGGAGGATCAACACTTTGTGCCCTTCAATGTTTGGAAAAGCTATA 
               
               
                   
               
               
                 TGGAGATAAATGGAATTCTTTTACCATCTTATTAATTCACTCTGGTGGCT 
               
               
                   
               
               
                 ACAGTCAACGACTTCCAAATGCAAGTGCTCTGGGAAAAATTTTCACTGCT 
               
               
                   
               
               
                 TTACCTCTTGGTAACCCCATTTATCAGATGCTAGAATTAAAGCTAGCCAT 
               
               
                   
               
               
                 GTACATTGATTTCCCCTTAAATATGAATCCTGGAATTCTGGTTACCTGTG 
               
               
                   
               
               
                 CAGATGATATTGAACTTTATAGTATTGGAGAATTTGAGTTTATTAGGTTT 
               
               
                   
               
               
                 GACAAACCTGGCTTTACTGCTTTAGCTCATCCTTCTAGTTTGACGATAGG 
               
               
                   
               
               
                 TACCACACATGGAGTATTTGTCTTAGATCCTTTTGATGATTTAAAACATA 
               
               
                   
               
               
                 GAGACCTTGAATACAGGTCTTGCCATCGTTTCCTTCATAAGCCCAGCATA 
               
               
                   
               
               
                 GAAAAGATGTATCAGTTTAATGCTGTGTGTAGACCTGGAAATTTTTGTCA 
               
               
                   
               
               
                 ACAGGACTTTGCTGGGGGTGACATTGCCGATCTTAAATTAGACTCTGACT 
               
               
                   
               
               
                 ATGTCTACACAGATAGCCTATTTTATATGGATCATAAATCAGCAAAAATG 
               
               
                   
               
               
                 TTACTTGCTTTTTATGAAAAAATAGGCACACTGAGCTGTGAAATAGATGC 
               
               
                   
               
               
                 CTATGGTGACTTTCTGCAGGCTTTGGGACCTGGAGCAACTGTGGAGTACA 
               
               
                   
               
               
                 CCAGAAACACATCACATGTCATTAAAGAAGAGTCAGAGTTGGTAGAAATG 
               
               
                   
               
               
                 AGGCAGAGAATATTTCATCTTCTTAAAGGAACATCACTAAATGTTGTTGT 
               
               
                   
               
               
                 TCTTAATAACTCCAAATTTTATCACATTGGAACAACCGAAGAATATTTGT 
               
               
                   
               
               
                 TTTACTTTACCTCAGATAACAGTTTAAAGTCAGAGCTCGGCTTACAGTCC 
               
               
                   
               
               
                 ATAACTTTTAGTATCTTTCCAGATATACCAGAATGCTCTGGCAAAACATC 
               
               
                   
               
               
                 CTGTATCATTCAAAGCATACTGGATTCAAGATGTTCTGTGGCACCTGGCT 
               
               
                   
               
               
                 CAGTTGTGGAGTATTCCAGATTGGGGCCTGATGTTTCAGTTGGGGAAAAC 
               
               
                   
               
               
                 TGCATTATTAGTGGTTCTTACATCCTAACAAAAGCTGCCCTCCCCGCACA 
               
               
                   
               
               
                 TTCTTTTGTATGTTCCTTAAGCTTAAAGATGAATAGATGCTTAAAGTATG 
               
               
                   
               
               
                 CAACTATGGCATTTGGAGTGCAAGACAACTTGAAAAAGAGTGTGAAAACA 
               
               
                   
               
               
                 TTGTCAGATATAAAGTTACTTCAATTCTTTGGAGTCTGTTTCCTGTCATG 
               
               
                   
               
               
                 CTTAGATGTTTGGAATCTTAAAGTTACAGAGGAACTGTTCTCTGGTAACA 
               
               
                   
               
               
                 AGACATGTCTGAGTTTGTGGACTGCACGCATTTTCCCAGTTTGTTCTTCT 
               
               
                   
               
               
                 TTGAGTGACTCAGTTATAACATCCCTAAAGATGTTAAATGCTGTTAAGAA 
               
               
                   
               
               
                 CAAGTCAGCATTCAGCCTGAATAGCTATAAGTTGCTGTCCATTGAAGAAA 
               
               
                   
               
               
                 TGCTTATCTACAAAGATGTAGAAGATATGATAACTTACAGGGAACAAATT 
               
               
                   
               
               
                 TTTCTAGAAATCAGTTTAAAAAGCAGTTTGATGTAG 
               
               
                   
               
               
                 GenBank Accession No. AF017445 (GenBank version 
               
               
                   
               
               
                 dated 12-NOV-1998) 
               
               
                   
               
               
                 Protein sequence of mouse GDP fucose pyrophos- 
               
               
                 phorylase (fucose-1-phosphate guanylyltransferase) 
               
               
                 (SEQ ID NO: 19) 
               
               
                 MASLREATLRKLRRFSELRGKPVAAGEFWDVVAITAADEKQELAYKQQLS 
               
               
                   
               
               
                 EKLKKRELPLGVQYHVFPDPAGTKIGNGGSTLCSLECLESLCGDKWNSLK 
               
               
                   
               
               
                 VLLIHSGGYSQRLPNASALGKIFTALPLGEPIYQMLELKLAMYVDFPSNM 
               
               
                   
               
               
                 RPGVLVTCADDIELYSVGDSEYIAFDQPGFTALAHPSSLAVGTTHGVFVL 
               
               
                   
               
               
                 HSDSSLQHGDLEYRQCYQFLHKPTIENMHRFNAVHRQRSFGQQNLSGGDT 
               
               
                   
               
               
                 DCLPLHTEYVYTDSLFYMDHKSAKKLLDFYKSEGPLNCEIDAYGDFLQAL 
               
               
                   
               
               
                 GPGATAEYTRNTSHVTKEESQLLDMRQKIFHLLKGTPLNVVVLNNSRFYH 
               
               
                   
               
               
                 IGTLQEYLLHFTSDSALKTELGLQSIAFSVSPSVPERSSGTACVIHSIVD 
               
               
                   
               
               
                 SGCCVAPGSVVEYSRLGPEVSIGENCIISSSVIAKTVVPAYSFLCSLSVK 
               
               
                   
               
               
                 INGHLKYSTMVFGMQDNLKNSVKTLEDIKALQFFGVCFLSCLDIWNLKAT 
               
               
                   
               
               
                 EKLFSGNKMNLSLWTACIFPVCSSLSESATASLGMLSAVRNHSPFNLSDF 
               
               
                   
               
               
                 NLLSIQEMLVYKDVQDMLAYREHIFLEISSNKNQSDLEKS 
               
               
                   
               
               
                 GenBank Accession No. NP_083606 (GenBank version 
               
               
                   
               
               
                 dated 10-FEB-2008) 
               
               
                   
               
               
                 mRNA sequence of mouse GDP fucose pyrophos- 
               
               
                 phorylase (fucose-1-phosphate guanylyltransferase) 
               
               
                 (SEQ ID NO: 20) 
               
               
                 agtgtgctcccggaagtcggccATGGCGTCTCTCCGCGAAGCCACCCTGC 
               
               
                   
               
               
                 GGAAACTGCGCAGATTTTCTGAGCTGAGAGGCAAACCCGTGGCAGCTGGA 
               
               
                   
               
               
                 GAATTCTGGGATGTGGTTGCAATAACAGCAGCTGATGAAAAGCAGGAGCT 
               
               
                   
               
               
                 CGCTTACAAGCAACAGTTGTCCGAGAAGCTGAAGAAAAGGGAATTGCCTC 
               
               
                   
               
               
                 TTGGAGTTCAATACCATGTTTTTCCAGATCCTGCTGGGACCAAAATTGGA 
               
               
                   
               
               
                 AATGGAGGATCAACACTTTGTTCCCTTGAGTGTTTGGAAAGCCTCTGTGG 
               
               
                   
               
               
                 AGACAAATGGAATTCTCTGAAGGTCCTGCTAATCCACTCTGGTGGCTACA 
               
               
                   
               
               
                 GCCAACGCCTTCCCAATGCGAGTGCTTTAGGAAAGATCTTCACAGCCTTA 
               
               
                   
               
               
                 CCACTTGGTGAACCCATTTATCAGATGTTGGAGTTAAAACTAGCCATGTA 
               
               
                   
               
               
                 CGTGGATTTCCCCTCAAACATGAGGCCTGGAGTCTTGGTCACCTGTGCAG 
               
               
                   
               
               
                 ATGATATCGAACTCTACAGTGTTGGGGACAGTGAGTACATTGCCTTTGAC 
               
               
                   
               
               
                 CAGCCTGGCTTTACTGCCTTAGCCCATCCGTCTAGTCTGGCTGTAGGCAC 
               
               
                   
               
               
                 TACTCATGGAGTATTTGTCTTGCACTCTGACAGTTCCTTACAACATGGTG 
               
               
                   
               
               
                 ACCTTGAGTACAGGCAATGCTACCAATTCCTCCACAAGCCCACCATTGAA 
               
               
                   
               
               
                 AACATGCACCGCTTTAATGCTGTGCATAGACAACGAAGCTTTGGTCAACA 
               
               
                   
               
               
                 GAACTTGTCTGGAGGTGACACTGACTGTCTTCCATTGCACACTGAGTATG 
               
               
                   
               
               
                 TCTACACAGATAGCCTGTTTTACATGGATCACAAATCAGCCAAAAAGTTA 
               
               
                   
               
               
                 CTTGATTTCTATAAAAGTGAAGGCCCACTGAACTGTGAAATAGATGCCTA 
               
               
                   
               
               
                 TGGAGACTTTCTTCAGGCACTGGGGCCTGGAGCAACTGCAGAGTACACCA 
               
               
                   
               
               
                 GGAACACATCTCATGTCACTAAAGAAGAGTCCCAGTTGTTGGACATGAGG 
               
               
                   
               
               
                 CAGAAAATATTCCACCTCCTCAAGGGAACACCACTGAATGTTGTTGTTCT 
               
               
                   
               
               
                 TAATAACTCCAGATTTTATCACATTGGAACACTGCAAGAGTATCTGCTTC 
               
               
                   
               
               
                 ATTTCACCTCTGATAGTGCATTAAAGACGGAGCTGGGCTTACAATCCATA 
               
               
                   
               
               
                 GCTTTCAGTGTCTCTCCAAGTGTTCCTGAGCGCTCCAGTGGAACAGCCTG 
               
               
                   
               
               
                 TGTCATTCACAGTATAGTGGATTCAGGATGCTGTGTGGCCCCTGGCTCAG 
               
               
                   
               
               
                 TGGTAGAGTATTCTAGATTGGGGCCTGAGGTGTCCATCGGGGAAAACTGC 
               
               
                   
               
               
                 ATTATCAGCAGTTCTGTCATAGCAAAAACTGTTGTGCCAGCATATTCTTT 
               
               
                   
               
               
                 TTTGTGTTCTTTAAGTGTGAAGATAAATGGACACTTAAAATATTCTACTA 
               
               
                   
               
               
                 TGGTGTTTGGCATGCAAGACAACTTGAAGAACAGTGTTAAAACACTGGAA 
               
               
                   
               
               
                 GACATAAAGGCACTTCAGTTCTTTGGAGTCTGTTTTCTGTCTTGTTTAGA 
               
               
                   
               
               
                 CATTTGGAATCTTAAAGCTACAGAGAAACTATTCTCTGGAAATAAGATGA 
               
               
                   
               
               
                 ATCTGAGCCTGTGGACTGCATGCATTTTCCCTGTCTGTTCATCTCTGAGT 
               
               
                   
               
               
                 GAGTCGGCTACAGCATCCCTTGGGATGTTAAGCGCTGTAAGGAACCATTC 
               
               
                   
               
               
                 ACCATTCAACCTAAGTGACTTTAACCTTTTGTCCATCCAGGAAATGCTTG 
               
               
                   
               
               
                 TCTACAAAGATGTACAAGACATGCTAGCTTATAGGGAACACATTTTTCTA 
               
               
                   
               
               
                 GAAATTAGTTCAAATAAAAATCAATCTGATTTAGAGAAATCTTGAatata 
               
               
                   
               
               
                 ttttggccataaacaaaattgcaaatacaggcattttctatagacctctg 
               
               
                   
               
               
                 acatttttgtttgttttaataaagtaatataataaaaattatgttaatat 
               
               
                   
               
               
                 aactgttgtagcttggtaatgagaatggtacaactgaccacttctgctag 
               
               
                   
               
               
                 aagtacgttccaggactagagtcaggaaaggtcggctgttttagatgttt 
               
               
                   
               
               
                 acaccatcttacaattgtgctctttggtaaagatccatttatgggacact 
               
               
                   
               
               
                 gtttcattcacaaaataaatatttctgttttataggatgattttctaaac 
               
               
                   
               
               
                 ataacatatctttaaagcttttctatcttcttttgaaatttggaccaata 
               
               
                   
               
               
                 aaattctaggtgatatggaggattgtattgctcaacttctcatagtgaga 
               
               
                   
               
               
                 caacacgtaacaaaacattgttataaattcttagaagaaatgtcattatt 
               
               
                   
               
               
                 tgaggttttctttgaggactttgttctagttttattttatgtgtataaat 
               
               
                   
               
               
                 gtgttacctgcatgtatgcatgtgcaccacttgcctgcggcacccataga 
               
               
                   
               
               
                 ggctagaacagctgttctcaacatttgggttgggaccttttgtgggctca 
               
               
                   
               
               
                 aacaatcctttgaggggtaacctaagtccattggaaaacaaaatatttac 
               
               
                   
               
               
                 attatgattcataacagtagggaaattacagttaagtagcaacaaaaata 
               
               
                   
               
               
                 attttatatttggggtcactacagcatggggactgtattgaaaggatagc 
               
               
                   
               
               
                 agcatcaggaaggttaaaaactgccggtctagaagaaagcattgggtctc 
               
               
                   
               
               
                 ttggaactagagttatagatgcttagaacctccgtgttgcttctgtaagt 
               
               
                   
               
               
                 caacctccttagtcctatgaaagtgctatataatgatgtttgtgcctcat 
               
               
                   
               
               
                 tggtcttgccaaaatgatataaaagtatgtatggatgattttgttcttat 
               
               
                   
               
               
                 acactagaacatgtgttgccatatcttataaactatgtctactgatatat 
               
               
                   
               
               
                 tacactggtagctatgtacacacagaactcagttgtctgctcaggaggtg 
               
               
                   
               
               
                 gtagggatagttgagagccagtactcactcactatggaccttacttaatc 
               
               
                   
               
               
                 ctctcctagttaatccttctccaaatctcttaacttgacagtggacattt 
               
               
                   
               
               
                 gccttgcatcattggtggtagtgatgctgtgaacaaacaataggcccaaa 
               
               
                   
               
               
                 gagaggaaattcaaataggcaatctgaagaactactcaaatcataaacaa 
               
               
                   
               
               
                 ctgcagggaaatgaaatgggtggaattcctggttatgcgtacctattatg 
               
               
                   
               
               
                 aaataaacacattagtggaatgtccttaggttgaactgtaatagagttaa 
               
               
                   
               
               
                 attttatcatacttgtgtttaaaataccttaagtacattgtaatatctgc 
               
               
                   
               
               
                 tgtggcaactttaattctgtgtaagttttcataaaaatatatgataaaca 
               
               
                   
               
               
                 agatatctgtcaaaactcctttatattatttatataagaatatttgcctt 
               
               
                   
               
               
                 tttgaggtactagataataaagcaaagaatgtacgatactatatgacaat 
               
               
                   
               
               
                 tattggtaaagttacagagaattcaatggatgttaaatgttattaaatac 
               
               
                   
               
               
                 tcaagactaaagtcctatcaacgatgagaattatgatttcatgttccaag 
               
               
                   
               
               
                 aaaaaaatatcattaataaagaataccatcacttccttgtaaaaaaaaaa 
               
               
                   
               
               
                 aaaaaaaaaaaaaaaaaaaaaaaaaa 
               
               
                   
               
               
                 GenBank Accession No. NM_029330 (GenBank version 
               
               
                   
               
               
                 dated 10-FEB-2008) 
               
               
                   
               
               
                 Protein sequence of rat GDP fucose pyrophos- 
               
               
                 phorylase (fucose-1-phosphate guanylyltransferase) 
               
               
                 (SEQ ID NO: 21) 
               
               
                 METLREATLRKLRRFSELRGKPVAAGEFWDVVAITAADEKQELAYKQQLS 
               
               
                   
               
               
                 EKLRRKELPLGVQYHVFPDPAGTKIGNGGSTLCSLQCLKSLYGDEWNSFK 
               
               
                   
               
               
                 VLLIHSGGYSQRLPNASALGKIFTALPLGEPIYQMLELKLAMYVDFPSHM 
               
               
                   
               
               
                 KPGVLVTCADDIELYSVGDCQYIAFDQPGFTALAHPSSLAVGTTHGVFVL 
               
               
                   
               
               
                 HSASSLQHGDLQYRQCHRFLHKPTIENMHQFNAVQRQGSFAQQDFPGGDT 
               
               
                   
               
               
                 ACLPLHTEYVYTDSLFYMDHKSAKKLLDFYKNVNQLNCEIDAYGDFLQAL 
               
               
                   
               
               
                 GPGATAEYTRNTSHVTKEDSQLLDMRQKIFHLLKGTPLNVVVLNNSRFYH 
               
               
                   
               
               
                 IGTTQEYLLHFTSDSTLRSRARLTVHSFQVSLQVSLNPPMKQPVSFTVYW 
               
               
                   
               
               
                 DSGCCVAPGSVVEYSRLGPEVSIGENCIVSSSVLANTAVPAYSFVCSLSV 
               
               
                   
               
               
                 RTNGLLEYSTMVFSVQDNLKGSVKTLEDIKALQFFGVCFLSCLDIWNLKA 
               
               
                   
               
               
                 TEKLFSGSKRNLSLWTARIFPVCPSLSESVTASLGMLSAVRSHSPFSLSN 
               
               
                   
               
               
                 FKLMSIQEMLVYKDVQDMLAYREQIFLEINSNKKQSDLEKS 
               
               
                   
               
               
                 GenBank Accession No. NP_955788 (GenBank version 
               
               
                   
               
               
                 dated 11-FEB-2008) 
               
               
                   
               
               
                 Protein sequence of rat GDP fucose pyrophos- 
               
               
                 phorylase (fucose-1-phosphate guanylyltransferase) 
               
               
                 (SEQ ID NO: 22) 
               
               
                 ATGGAGACTCTCCGGGAAGCCACCCTGCGGAAACTGCGCAGATTTTCGGA 
               
               
                   
               
               
                 GCTGAGAGGCAAACCTGTGGCAGCTGGAGAATTCTGGGATGTGGTTGCGA 
               
               
                   
               
               
                 TAACAGCAGCCGATGAAAAGCAGGAGCTCGCTTACAAGCAGCAGTTGTCA 
               
               
                   
               
               
                 GAAAAGCTGAGAAGAAAGGAATTGCCTCTTGGAGTTCAATACCATGTTTT 
               
               
                   
               
               
                 TCCTGATCCTGCTGGGACCAAAATTGGAAATGGAGGATCGACACTTTGTT 
               
               
                   
               
               
                 CCCTTCAGTGCCTAAAAAGCCTCTATGGAGATGAATGGAATTCTTTCAAG 
               
               
                   
               
               
                 GTCCTGTTAATTCACTCCGGTGGCTACAGTCAACGCCTTCCCAATGCAAG 
               
               
                   
               
               
                 TGCTTTAGGAAAGATCTTCACAGCCTTACCACTTGGTGAACCCATCTATC 
               
               
                   
               
               
                 AGATGTTGGAGTTAAAACTAGCCATGTACGTGGATTTCCCCTCACACATG 
               
               
                   
               
               
                 AAGCCTGGAGTCTTGGTCACCTGTGCAGATGACATTGAACTGTACAGTGT 
               
               
                   
               
               
                 TGGGGACTGTCAGTACATTGCCTTTGACCAGCCTGGCTTTACTGCCTTAG 
               
               
                   
               
               
                 CCCATCCTTCCAGTCTGGCTGTAGGCACCACACACGGAGTATTTGTCTTG 
               
               
                   
               
               
                 CACTCTGCCAGTTCCTTACAACATGGTGACCTTCAGTACAGACAATGCCA 
               
               
                   
               
               
                 CCGTTTCCTCCACAAGCCCACCATTGAAAACATGCATCAGTTTAATGCTG 
               
               
                   
               
               
                 TGCAAAGACAAGGAAGCTTTGCTCAACAGGACTTCCCTGGAGGTGACACC 
               
               
                   
               
               
                 GCGTGTCTTCCATTGCACACTGAGTATGTCTACACAGATAGCCTGTTTTA 
               
               
                   
               
               
                 CATGGACCACAAATCGGCCAAAAAGTTACTTGATTTCTATAAAAATGTAA 
               
               
                   
               
               
                 ACCAACTGAACTGTGAAATAGATGCCTATGGTGACTTTCTGCAGGCACTG 
               
               
                   
               
               
                 GGGCCTGGAGCAACTGCAGAGTATACCAGGAACACATCACATGTCACTAA 
               
               
                   
               
               
                 AGAAGACTCCCAGTTGTTGGACATGAGGCAGAAAATATTCCACCTCCTCA 
               
               
                   
               
               
                 AGGGGACACCACTGAATGTTGTTGTTCTTAATAACTCCAGATTTTATCAC 
               
               
                   
               
               
                 ATTGGAACAACACAAGAATATCTGCTTCATTTCACGTCTGATAGTACGTT 
               
               
                   
               
               
                 AAGGTCAAGAGCTAGGCTTACAGTCCATAGCTTTCAAGTGTCTCTCCAAG 
               
               
                   
               
               
                 TATCCCTGAATCCTCCAATGAAACAGCCTGTATCATTCACAGTATACTGG 
               
               
                   
               
               
                 GATTCAGGATGCTGTGTGGCACCTGGCTCAGTTGTAGAGTATTCTAGACT 
               
               
                   
               
               
                 GGGGCCTGAGGTGTCCATTGGGGAAAACTGCATTGTCAGCAGCTCTGTCC 
               
               
                   
               
               
                 TAGCAAACACTGCTGTGCCGGCATATTCTTTTGTGTGTTCTCTAAGTGTG 
               
               
                   
               
               
                 AGGACAAATGGACTCTTGGAATATTCTACCATGGTGTTTAGTGTGCAGGA 
               
               
                   
               
               
                 CAACTTGAAAGGCAGTGTTAAAACCCTGGAAGATATAAAGGCACTTCAGT 
               
               
                   
               
               
                 TCTTTGGAGTCTGTTTCTTGTCTTGTTTAGACATCTGGAACCTTAAAGCT 
               
               
                   
               
               
                 ACAGAGAAACTGTTCTCTGGAAGTAAGAGGAACCTGAGCCTGTGGACTGC 
               
               
                   
               
               
                 ACGGATTTTCCCTGTCTGTCCTTCTCTGAGTGAGTCAGTTACAGCATCCC 
               
               
                   
               
               
                 TTGGGATGTTAAGTGCTGTAAGGAGCCATTCACCATTCAGCCTAAGCAAC 
               
               
                   
               
               
                 TTTAAGCTGATGTCCATCCAGGAAATGCTTGTCTACAAAGATGTACAAGA 
               
               
                   
               
               
                 CATGCTAGCTTATAGGGAGCAAATTTTTCTAGAAATTAATTCAAATAAAA 
               
               
                   
               
               
                 AACAATCTGATTTAGAGAAATCTTAA 
               
               
                   
               
               
                 GenBank Accession No. NM_199494 (GenBank version 
               
               
                   
               
               
                 dated 11-FEB-2008) 
               
               
                   
               
               
                 Protein sequence of human fucose kinase 
               
               
                 (fucokinase) 
               
               
                 (SEQ ID NO: 23) 
               
               
                 MEQPKGVDWTVIILTCQYKDSVQVFQRELEVRQKREQIPAGTLLLAVEDP 
               
               
                   
               
               
                 EKRVGSGGATLNALLVAAEHLSARAGFTVVTSDVLHSAWILILHMGRDFP 
               
               
                   
               
               
                 FDDCGRAFTCLPVENPEAPVEALVCNLDCLLDIMTYRLGPGSPPGVWVCS 
               
               
                   
               
               
                 TDMLLSVPANPGISWDSFRGARVIALPGSPAYAQNHGVYLTDPQGLVLDI 
               
               
                   
               
               
                 YYQGTEAEIQRCVRPDGRVPLVSGVVFFSVETAERLLATHVSPPLDACTY 
               
               
                   
               
               
                 LGLDSGARPVQLSLFFDILHCMAENVTREDFLVGRPPELGQGDADVAGYL 
               
               
                   
               
               
                 QSARAQLWRELRDQPLTMAYVSSGSYSYMTSSASEFLLSLTLPGAPGAQI 
               
               
                   
               
               
                 VHSQVEEQQLLAAGSSVVSCLLEGPVQLGPGSVLQHCHLQGPIHIGAGCL 
               
               
                   
               
               
                 VTGLDTAHSKALHGRELRDLVLQGHHTRLHGSPGHAFTLVGRLDSWERQG 
               
               
                   
               
               
                 AGTYLNVPWSEFFKRTGVRAWDLWDPETLPAEYCLPSARLFPVLHPSREL 
               
               
                   
               
               
                 GPQDLLWMLDHQEDGGEALRAWRASWRLSWEQLQPCLDRAATLASRRDLF 
               
               
                   
               
               
                 FRQALHKARHVLEARQDLSLRPLIWAAVREGCPGPLLATLDQVAAGAGDP 
               
               
                   
               
               
                 GVAARALACVADVLGCMAEGRGGLRSGPAANPEWMRPFSYLECGDLAAGV 
               
               
                   
               
               
                 EALAQERDKWLSRPALLVRAARHYEGAGQILIRQAVMSAQHFVSTEQVEL 
               
               
                   
               
               
                 PGPGQWVVAECPARVDFSGGWSDTPPLAYELGGAVLGLAVRVDGRRPIGA 
               
               
                   
               
               
                 RARRIPEPELWLAVGPRQDEMTVKIVCRCLADLRDYCQPHAPGALLKAAF 
               
               
                   
               
               
                 ICAGIVHVHSELQLSEQLLRTFGGGFELHTWSELPHGSGLGTSSILAGTA 
               
               
                   
               
               
                 LAALQRAAGRVVGTEALIHAVLHLEQVLTTGGGWQDQVGGLMPGIKVGRS 
               
               
                   
               
               
                 RAQLPLKVEVEEVTVPEGFVQKLNDHLLLVYTGKTRLARNLLQDVLRSWY 
               
               
                   
               
               
                 ARLPAVVQNAHSLVRQTEECAEGFRQGSLPLLGQCLTSYWEQKKLMAPGC 
               
               
                   
               
               
                 EPLTVRRMMDVLAPHVHGQSLAGAGGGGFLYLLTKEPQQKEALEAVLAKT 
               
               
                   
               
               
                 EGLGNYSIHLVEVDTQGLSLKLLGTEASTCCPFP 
               
               
                   
               
               
                 GenBank Accession No. NP_659496 (GenBank version 
               
               
                   
               
               
                 dated 22-OCT-2008) 
               
               
                   
               
               
                 mRNA sequence of human fucose kinase (fucokinase) 
               
               
                 (SEQ ID NO: 24) 
               
               
                 ATGGAGCAGCCGAAGGGAGTTGATTGGACAGTCATCATCCTGACCTGCCA 
               
               
                   
               
               
                 GTACAAGGACAGTGTCCAGGTCTTTCAGAGAGAACTGGAAGTGCGGCAGA 
               
               
                   
               
               
                 AGCGGGAGCAGATCCCTGCTGGGACGCTGTTACTGGCCGTGGAGGACCCA 
               
               
                   
               
               
                 GAGAAGCGTGTGGGCAGCGGAGGAGCCACCCTCAACGCCCTGCTGGTGGC 
               
               
                   
               
               
                 TGCTGAACACCTGAGTGCCCGGGCAGGCTTCACTGTGGTCACATCCGATG 
               
               
                   
               
               
                 TCCTGCACTCGGCCTGGATCCTCATTCTGCACATGGGTCGAGACTTCCCC 
               
               
                   
               
               
                 TTTGATGACTGTGGCAGGGCTTTCACCTGCCTCCCCGTGGAGAACCCCGA 
               
               
                   
               
               
                 GGCCCCCGTGGAAGCCTTGGTCTGCAACCTGGACTGCCTGCTGGACATCA 
               
               
                   
               
               
                 TGACCTATCGGCTGGGCCCGGGCTCCCCGCCAGGCGTGTGGGTCTGCAGC 
               
               
                   
               
               
                 ACCGACATGCTGCTGTCTGTTCCTGCAAATCCTGGTATCAGCTGGGACAG 
               
               
                   
               
               
                 CTTCCGGGGAGCCAGAGTGATCGCCCTCCCAGGGAGCCCGGCCTACGCTC 
               
               
                   
               
               
                 AGAATCATGGCGTCTACCTAACTGACCCCCAGGGCCTTGTTTTGGACATT 
               
               
                   
               
               
                 TACTACCAGGGCACTGAGGCAGAGATTCAGCGGTGTGTCAGGCCTGATGG 
               
               
                   
               
               
                 GCGGGTGCCACTGGTCTCTGGGGTTGTCTTCTTCTCTGTGGAGACTGCCG 
               
               
                   
               
               
                 AGCGCCTCCTAGCCACCCACGTGAGCCCGCCCCTGGATGCCTGCACCTAC 
               
               
                   
               
               
                 CTAGGCTTGGACTCCGGAGCCCGGCCTGTCCAGCTGTCTCTGTTTTTTGA 
               
               
                   
               
               
                 CATTCTCCACTGCATGGCTGAGAACGTGACCAGGGAGGACTTCCTGGTGG 
               
               
                   
               
               
                 GGAGGCCCCCAGAGTTGGGGCAAGGCGATGCAGATGTAGCGGGTTATCTG 
               
               
                   
               
               
                 CAGAGCGCCCGGGCCCAGCTGTGGAGGGAGCTTCGCGATCAGCCCCTTAC 
               
               
                   
               
               
                 CATGGCCTATGTCTCCAGCGGCAGCTACAGCTACATGACCTCCTCAGCCA 
               
               
                   
               
               
                 GTGAGTTCCTGCTCAGCCTCACACTCCCCGGGGCTCCTGGGGCCCAGATT 
               
               
                   
               
               
                 GTGCACTCCCAGGTGGAGGAGCAGCAGCTTCTGGCGGCCGGGAGCTCTGT 
               
               
                   
               
               
                 GGTCAGCTGCCTGCTGGAGGGCCCTGTCCAGCTGGGTCCTGGGAGCGTCC 
               
               
                   
               
               
                 TGCAGCACTGCCACCTGCAGGGCCCCATTCACATAGGCGCTGGCTGCTTG 
               
               
                   
               
               
                 GTGACTGGCCTGGATACAGCCCACTCCAAGGCCCTGCATGGCCGGGAGCT 
               
               
                   
               
               
                 GCGTGACCTTGTCCTGCAGGGACACCACACGCGGCTACACGGCTCCCCGG 
               
               
                   
               
               
                 GCCACGCCTTCACCCTCGTTGGCCGTCTGGACAGCTGGGAGAGACAGGGG 
               
               
                   
               
               
                 GCAGGCACATATCTCAACGTGCCCTGGAGTGAATTCTTCAAGAGGACAGG 
               
               
                   
               
               
                 TGTTCGAGCCTGGGACCTGTGGGACCCTGAGACGCTGCCCGCAGAGTACT 
               
               
                   
               
               
                 GCCTTCCCAGCGCCCGCCTCTTTCCTGTGCTCCACCCCTCGAGGGAGCTG 
               
               
                   
               
               
                 GGACCCCAGGACCTGCTGTGGATGCTGGACCACCAGGAGGATGGGGGCGA 
               
               
                   
               
               
                 GGCCCTGCGAGCCTGGCGGGCCTCCTGGCGCCTGTCCTGGGAGCAGCTGC 
               
               
                   
               
               
                 AGCCGTGCCTGGATCGGGCTGCCACGCTGGCCTCTCGCCGGGACCTGTTC 
               
               
                   
               
               
                 TTCCGCCAGGCCCTGCATAAGGCGCGGCACGTGCTGGAGGCCCGGCAGGA 
               
               
                   
               
               
                 CCTCAGCCTGCGCCCGCTGATCTGGGCTGCTGTCCGCGAGGGCTGCCCCG 
               
               
                   
               
               
                 GGCCCCTGCTGGCCACGCTGGACCAGGTTGCAGCTGGGGCAGGAGACCCT 
               
               
                   
               
               
                 GGTGTGGCGGCACGGGCACTGGCCTGTGTGGCGGACGTCCTGGGCTGCAT 
               
               
                   
               
               
                 GGCAGAGGGCCGTGGGGGCTTGCGGAGCGGGCCAGCTGCCAACCCTGAGT 
               
               
                   
               
               
                 GGATGCGGCCCTTCTCATACCTGGAGTGTGGAGACCTGGCAGCGGGCGTG 
               
               
                   
               
               
                 GAGGCGCTTGCCCAGGAGAGGGACAAGTGGCTAAGCAGGCCAGCCTTGCT 
               
               
                   
               
               
                 GGTGCGAGCGGCCCGCCACTATGAGGGGGCTGGTCAGATCCTGATCCGCC 
               
               
                   
               
               
                 AGGCTGTGATGTCAGCCCAGCACTTTGTCTCCACAGAGCAGGTGGAACTG 
               
               
                   
               
               
                 CCGGGACCTGGGCAGTGGGTGGTGGCTGAGTGCCCGGCCCGTGTGGATTT 
               
               
                   
               
               
                 CTCTGGGGGCTGGAGTGACACGCCACCCCTTGCCTATGAGCTTGGCGGGG 
               
               
                   
               
               
                 CTGTGCTGGGCCTGGCTGTGCGAGTGGACGGCCGCCGGCCCATCGGAGCC 
               
               
                   
               
               
                 AGGGCACGCCGCATCCCGGAGCCTGAGCTGTGGCTGGCGGTGGGGCCTCG 
               
               
                   
               
               
                 GCAGGATGAGATGACTGTGAAGATAGTGTGCCGGTGCCTGGCTGACCTGC 
               
               
                   
               
               
                 GGGACTACTGCCAGCCTCATGCCCCAGGGGCCCTGCTGAAGGCGGCCTTC 
               
               
                   
               
               
                 ATCTGTGCAGGGATCGTGCATGTCCACTCGGAACTCCAGCTGAGTGAGCA 
               
               
                   
               
               
                 GCTGCTCCGCACCTTCGGGGGCGGCTTTGAGCTGCACACCTGGTCTGAGC 
               
               
                   
               
               
                 TGCCCCACGGCTCTGGCCTGGGCACCAGCAGCATCCTGGCAGGCACTGCC 
               
               
                   
               
               
                 CTGGCTGCCTTGCAGCGAGCCGCAGGCCGGGTGGTGGGCACGGAAGCCCT 
               
               
                   
               
               
                 GATCCACGCAGTGCTGCACCTGGAGCAGGTGCTCACCACTGGAGGTGGCT 
               
               
                   
               
               
                 GGCAGGACCAAGTAGGTGGCCTAATGCCTGGCATCAAGGTGGGGCGCTCC 
               
               
                   
               
               
                 CGGGCTCAGCTGCCACTGAAGGTGGAGGTAGAAGAGGTCACGGTGCCTGA 
               
               
                   
               
               
                 GGGCTTTGTCCAGAAGCTCAATGACCACCTGCTCTTGGTGTACACTGGCA 
               
               
                   
               
               
                 AGACCCGCCTGGCTCGGAACCTGCTGCAGGATGTGCTGAGGAGCTGGTAT 
               
               
                   
               
               
                 GCCCGACTTCCTGCTGTGGTGCAGAATGCCCACAGCCTGGTACGGCAAAC 
               
               
                   
               
               
                 TGAGGAGTGTGCTGAAGGCTTCCGCCAAGGAAGCCTGCCTCTGCTGGGCC 
               
               
                   
               
               
                 AGTGCCTGACCTCGTACTGGGAGCAGAAGAAGCTCATGGCTCCAGGCTGT 
               
               
                   
               
               
                 GAGCCCCTGACTGTGCGGCGTATGATGGATGTCCTGGCCCCCCACGTGCA 
               
               
                   
               
               
                 TGGCCAGAGCCTGGCTGGGGCAGGCGGTGGAGGCTTTCTCTATCTGTTGA 
               
               
                   
               
               
                 CCAAGGAGCCACAGCAAAAGGAGGCCTTGGAGGCGGTGCTGGCCAAGACC 
               
               
                   
               
               
                 GAGGGCCTTGGGAATTACAGCATCCACCTGGTTGAAGTGGACACTCAGGG 
               
               
                   
               
               
                 CCTGAGCCTGAAGCTGCTGGGGACCGAGGCCTCAACCTGTTGCCCTTTCC 
               
               
                   
               
               
                 CATGA 
               
               
                   
               
               
                 GenBank Accession No. NM_145059 (GenBank version 
               
               
                   
               
               
                 dated 22-OCT-2008) 
               
               
                   
               
               
                 Protein sequence of mouse fucose kinase 
               
               
                 (fucokinase) 
               
               
                 (SEQ ID NO: 25) 
               
               
                 MEQSEGVNWTVIILTCQYKDSVQVFQRELEVRQRREQIPAGTMLLAVEDP 
               
               
                   
               
               
                 QTRVGSGGATLNALLVAAEHLSARAGFTVVTSDVLHSAWILILHMGRDFP 
               
               
                   
               
               
                 FDDCGRAFTCLPVENPQAPVEALVCNLDCLLDIMTHRLGPGSPPGVWVCS 
               
               
                   
               
               
                 TDMLLSVPPNPGISWDGFRGARVIAFPGSLAYALNHGVYLTDSQGLVLDI 
               
               
                   
               
               
                 YYQGTKAEIQRCVGPDGLVPLVSGVVFFSVETAEHLLATHVSPPLDACTY 
               
               
                   
               
               
                 MGLDSGAQPVQLSLFFDILLCMARNMSRENFLAGRPPELGQGDMDVASYL 
               
               
                   
               
               
                 KGARAQLWRELRDQPLTMVYVPDGGYSYMTTDATEFLHRLTMPGVAVAQI 
               
               
                   
               
               
                 VHSQVEEPQLLEATCSVVSCLLEGPVHLGPRSVLQHCHLRGPIRIGAGCF 
               
               
                   
               
               
                 VSGLDTAHSEALHGLELHDVILQGHHVRLHGSLSRVFTLAGRLDSWERQG 
               
               
                   
               
               
                 AGMYLNMSWNEFFKKTGIRDWDLWDPDTPPSDRCLLTARLFPVLHPTRAL 
               
               
                   
               
               
                 GPQDVLWMLHPRKHRGEALRAWRASWRLSWEQLQPCVDRAATLDFRRDLF 
               
               
                   
               
               
                 FCQALQKARHVLEARQDLCLRPLIRAAVGEGCSGPLLATLDKVAAGAEDP 
               
               
                   
               
               
                 GVAARALACVADVLGCMAEGRGGLRSGPAANPEWIQPFSYLECGDLMRGV 
               
               
                   
               
               
                 EALAQEREKWLTRPALLVRAARHYEGAEQILIRQAVMTARHFVSTQPVEL 
               
               
                   
               
               
                 PAPGQWVVTECPARVDFSGGWSDTPPIAYELGGAVLGLAVRVDGRRPIGA 
               
               
                   
               
               
                 KARRIPEPELWLAVGPRQDEMTMRIVCRSLDDLRDYCQPHAPGALLKAAF 
               
               
                   
               
               
                 ICAGIVHLHSELPLLEQLLHSFNGGFELHTWSELPHGSGLGTSSILAGAA 
               
               
                   
               
               
                 LAALQRAAGRAVGTEALIHAVLHLEQVLTTGGGWQDQVSGLMPGIKVGRS 
               
               
                   
               
               
                 RAQLPLKVEVEEITVPEGFVQKINDHLLLVYTGKTRLARNLLQDVLRNWY 
               
               
                   
               
               
                 ARLPVVVQNARRLVRQTEKCAEAFRQGNLPLLGQYLTSYWEQKKLMAPGC 
               
               
                   
               
               
                 EPLAVQRMMDVLAPYAYGQSLAGAGGGGFLYLLTKEPRQKETLEAVLAKA 
               
               
                   
               
               
                 EGLGNYSVHLVEVDPQGLSLQLLGHDTRLCGAGPSEVGTT 
               
               
                   
               
               
                 GenBank Accession No. NP_758487 (GenBank version 
               
               
                   
               
               
                 dated 05-AUG-2008) 
               
               
                   
               
               
                 mRNA sequence of mouse fucose kinase (fucokinase) 
               
               
                 (SEQ ID NO: 26) 
               
               
                 ATGGAGCAGTCAGAGGGAGTCAATTGGACTGTCATTATCCTGACATGCCA 
               
               
                   
               
               
                 GTACAAGGACAGTGTCCAGGTCTTTCAGAGAGAGCTGGAGGTAAGGCAG 
               
               
                   
               
               
                 AGACGGGAGCAGATTCCTGCGGGGACGATGTTACTGGCTGTGGAGGATCC 
               
               
                   
               
               
                 CCAGACTCGAGTCGGCAGCGGAGGAGCCACCCTCAACGCACTGCTGGTGG 
               
               
                   
               
               
                 CTGCTGAACACTTGAGTGCCCGAGCTGGCTTCACTGTGGTCACGTCCGAT 
               
               
                   
               
               
                 GTCCTGCACTCTGCCTGGATCCTCATCTTGCACATGGGCCGAGACTTCCC 
               
               
                   
               
               
                 CTTCGATGACTGTGGCAGGGCCTTCACTTGCCTCCCTGTGGAGAACCCAC 
               
               
                   
               
               
                 AGGCCCCTGTGGAGGCCTTGGTATGCAACCTGGACTGCCTGTTGGATATC 
               
               
                   
               
               
                 ATGACCCACCGGCTGGGTCCAGGTTCCCCACCAGGTGTGTGGGTCTGCAG 
               
               
                   
               
               
                 CACCGACATGCTTCTGTCTGTTCCTCCAAACCCTGGGATCAGTTGGGATG 
               
               
                   
               
               
                 GCTTCCGGGGAGCCAGAGTGATCGCCTTTCCTGGGAGCCTGGCCTATGCG 
               
               
                   
               
               
                 TTGAACCACGGTGTCTACCTCACTGACTCACAGGGCTTGGTTTTGGACAT 
               
               
                   
               
               
                 TTACTACCAGGGCACTAAGGCGGAGATACAACGTTGTGTCGGACCTGATG 
               
               
                   
               
               
                 GGCTGGTACCATTGGTCTCCGGGGTCGTCTTCTTCTCTGTGGAGACTGCT 
               
               
                   
               
               
                 GAGCACCTCCTAGCCACCCATGTGAGCCCACCGCTGGATGCCTGCACCTA 
               
               
                   
               
               
                 TATGGGCTTGGACTCTGGAGCCCAGCCTGTGCAGCTGTCTCTGTTTTTCG 
               
               
                   
               
               
                 ACATCCTGCTCTGCATGGCTCGGAATATGAGCAGGGAGAACTTCCTGGCT 
               
               
                   
               
               
                 GGGCGGCCCCCGGAGTTGGGGCAAGGTGACATGGATGTAGCAAGTTACCT 
               
               
                   
               
               
                 GAAGGGAGCCCGGGCCCAGCTGTGGAGGGAGCTTCGAGATCAGCCCCTCA 
               
               
                   
               
               
                 CAATGGTGTATGTCCCTGACGGCGGCTACAGCTACATGACGACTGATGCC 
               
               
                   
               
               
                 ACCGAGTTCCTGCACAGACTCACGATGCCTGGAGTAGCTGTGGCACAGAT 
               
               
                   
               
               
                 TGTTCACTCCCAGGTGGAGGAGCCACAGCTGCTAGAGGCTACGTGCTCGG 
               
               
                   
               
               
                 TGGTCAGCTGCCTGCTCGAGGGCCCTGTGCACCTGGGGCCTCGAAGTGTC 
               
               
                   
               
               
                 CTGCAGCACTGTCACCTGAGGGGCCCCATTCGCATCGGCGCTGGCTGCTT 
               
               
                   
               
               
                 TGTGAGTGGTCTGGATACAGCCCACTCGGAGGCACTGCATGGCCTGGAGC 
               
               
                   
               
               
                 TCCATGATGTCATCCTGCAGGGACACCATGTGCGGCTGCATGGCTCCCTG 
               
               
                   
               
               
                 AGCCGTGTATTTACTCTTGCTGGCCGTCTGGACAGCTGGGAAAGACAGGG 
               
               
                   
               
               
                 GGCAGGCATGTATCTCAACATGTCCTGGAATGAGTTCTTCAAGAAGACAG 
               
               
                   
               
               
                 GCATTCGAGACTGGGACCTGTGGGACCCAGATACACCCCCCTCAGATCGA 
               
               
                   
               
               
                 TGCCTCCTCACTGCCCGCCTTTTCCCTGTGCTCCACCCCACGAGGGCCCT 
               
               
                   
               
               
                 GGGGCCCCAGGATGTGCTGTGGATGCTGCACCCCCGCAAACACAGAGGTG 
               
               
                   
               
               
                 AGGCCCTTCGGGCCTGGCGAGCCTCCTGGCGTCTGTCCTGGGAGCAGCTG 
               
               
                   
               
               
                 CAACCTTGTGTGGACCGGGCTGCCACACTGGACTTCCGCCGAGATCTGTT 
               
               
                   
               
               
                 CTTCTGCCAGGCCTTGCAGAAGGCAAGGCATGTGTTAGAGGCGCGGCAGG 
               
               
                   
               
               
                 ACCTCTGCCTACGTCCACTGATCCGGGCCGCTGTCGGGGAAGGTTGCTCT 
               
               
                   
               
               
                 GGGCCCCTGCTGGCCACACTTGACAAGGTTGCAGCTGGGGCAGAAGATCC 
               
               
                   
               
               
                 TGGCGTGGCAGCCCGGGCTCTGGCTTGTGTGGCCGATGTGCTGGGCTGCA 
               
               
                   
               
               
                 TGGCAGAGGGCCGAGGAGGCTTGCGCAGTGGGCCAGCTGCCAACCCTGAG 
               
               
                   
               
               
                 TGGATTCAGCCTTTCTCATACTTGGAGTGTGGAGACCTGATGAGGGGTGT 
               
               
                   
               
               
                 GGAGGCGCTTGCCCAGGAGAGAGAGAAGTGGCTGACCAGGCCTGCCTTGC 
               
               
                   
               
               
                 TGGTTCGAGCTGCCCGCCATTACGAGGGGGCCGAGCAGATCCTGATCCGC 
               
               
                   
               
               
                 CAGGCTGTGATGACAGCCCGGCACTTCGTCTCCACCCAGCCCGTGGAGCT 
               
               
                   
               
               
                 GCCCGCACCCGGGCAGTGGGTGGTGACTGAGTGCCCAGCCCGTGTGGATT 
               
               
                   
               
               
                 TCTCTGGGGGCTGGAGTGACACACCGCCCATTGCCTATGAGCTTGGTGGA 
               
               
                   
               
               
                 GCAGTGTTGGGCCTGGCTGTGCGGGTGGATGGCCGCCGGCCCATCGGGGC 
               
               
                   
               
               
                 CAAAGCACGCCGCATCCCGGAGCCTGAGCTCTGGCTGGCAGTGGGACCTC 
               
               
                   
               
               
                 GGCAGGATGAGATGACCATGAGGATAGTGTGCCGGAGCCTGGATGACCTG 
               
               
                   
               
               
                 CGGGATTACTGCCAGCCTCATGCCCCAGGGGCCTTGCTGAAGGCAGCCTT 
               
               
                   
               
               
                 TATCTGTGCTGGCATTGTGCATCTCCACTCAGAGCTCCCTCTGCTTGAAC 
               
               
                   
               
               
                 AGTTGTTACACTCCTTTAATGGTGGCTTTGAGCTGCACACGTGGTCAGAG 
               
               
                   
               
               
                 CTGCCGCACGGCTCTGGTCTTGGCACCAGCAGCATCCTGGCAGGGGCTGC 
               
               
                   
               
               
                 CCTGGCTGCCTTACAGCGGGCTGCAGGCCGGGCAGTGGGCACGGAGGCTC 
               
               
                   
               
               
                 TCATCCACGCAGTGCTGCACCTGGAGCAGGTGCTCACCACAGGAGGTGGC 
               
               
                   
               
               
                 TGGCAGGACCAAGTCAGTGGCCTAATGCCTGGCATCAAAGTGGGGCGCTC 
               
               
                   
               
               
                 CCGGGCCCAGCTGCCCCTCAAGGTGGAGGTGGAGGAAATCACTGTGCCTG 
               
               
                   
               
               
                 AGGGCTTTGTCCAGAAGATCAATGACCATCTGCTCCTGGTTTATACCGGC 
               
               
                   
               
               
                 AAGACCCGATTGGCCCGGAATCTGCTGCAGGACGTGCTGAGGAACTGGTA 
               
               
                   
               
               
                 CGCTCGGTTGCCCGTTGTGGTACAGAATGCCCGCAGACTGGTGCGACAGA 
               
               
                   
               
               
                 CCGAGAAGTGCGCTGAAGCTTTCCGCCAAGGAAACCTGCCTCTGCTGGGA 
               
               
                   
               
               
                 CAGTACCTGACCTCATACTGGGAGCAGAAGAAGCTTATGGCCCCAGGCTG 
               
               
                   
               
               
                 CGAGCCGCTGGCCGTGCAGCGAATGATGGATGTCCTGGCCCCGTATGCGT 
               
               
                   
               
               
                 ATGGCCAAAGCCTGGCAGGGGCAGGTGGTGGGGGCTTTCTCTATCTATTG 
               
               
                   
               
               
                 ACCAAGGAACCCCGGCAGAAAGAGACTCTGGAAGCTGTCCTGGCCAAGGC 
               
               
                   
               
               
                 TGAGGGCCTTGGCAACTACAGTGTCCACCTGGTGGAAGTGGATCCTCAGG 
               
               
                   
               
               
                 GCCTGAGCCTGCAGCTGCTGGGACACGACACCCGTCTTTGTGGGGCCGGG 
               
               
                   
               
               
                 CCCTCTGAAGTGGGCACCACCTAG 
               
               
                   
               
               
                 GenBank Accession No. NM_172283 (GenBank version 
               
               
                   
               
               
                 dated 05-AUG-2008) 
               
               
                   
               
               
                 Protein sequence of rat fucose kinase (fucokinase) 
               
               
                 (SEQ ID NO: 27) 
               
               
                 MDQPKGVNWTVIILTCQYKDSVQVFQRELEVRQKREQIPAGTMLLAVEDP 
               
               
                   
               
               
                 QTRVGSGGATLNALLVAAEHLSARAGFTVVTSDVLHSAWILILHMGRDFP 
               
               
                   
               
               
                 FDDCGRAFTCLPVENPQAPVEALVCNLDCLLDIMTHRLGPGSPPGVWVCS 
               
               
                   
               
               
                 TDMLLSVPPNPGISWDGFRGTRVIAFPGSLAYALNHGVYLTDSQGVVLDI 
               
               
                   
               
               
                 YYQGTKAEIQRCVRPDGLVPLVSGVVFFSVETAEHLLATHVSPPLDACTY 
               
               
                   
               
               
                 MGLDSGAQPVQLSLFFDILLCMARNMSRENFVAGRPPEMGQGDPDVARYL 
               
               
                   
               
               
                 KGARAQLWRELRDQPLTMVYVPDGGYSYMTTDATEFLHRLTMPGVAVAQI 
               
               
                   
               
               
                 VHSQVEEPQLLEATCSVVSCLLEGPVHLGPRSVLQHCHLRGPIHIGAGCF 
               
               
                   
               
               
                 VSGLDTAHSEALHGLELHDLILQGHHIRLHGSQSRVFTLAGRLDSWERQG 
               
               
                   
               
               
                 AGMYLNMSWNEFFKKTGIRDWDLWDPDTPLSDRCLLSARLFPVLHPTRAL 
               
               
                   
               
               
                 GPQDVLWMLHPHKDRGEALRAWRASWRLSWEQLQPRLDRAATLDFRRDLF 
               
               
                   
               
               
                 FRQALQKARHVLEARQDLCLHPLIRAAVGEGCSGPLLATLDKVAAGAEDP 
               
               
                   
               
               
                 GVAARALACVADVLGCMAEGQGGLRSGPAANPEWIQPFSYLERGDLMRGV 
               
               
                   
               
               
                 EALAQEREKWLTRPALLVRAARHYEGAEQILIRQAVMTARHFVSTQPVEL 
               
               
                   
               
               
                 PAPGQWVVTECPARVDFSGGWSDTPPIAYELGGAVLGLAVRVDGRRPIGA 
               
               
                   
               
               
                 KARRILEPELWLAVGPRQDEMTVKIVCRSLDDLQDYCQPHAPGALLKAAF 
               
               
                   
               
               
                 ICADIVHVNSEVPLHEQLLRSFNGGFELHTWSELPHGSGLGTSSILAGAA 
               
               
                   
               
               
                 LAALQRAAGRTVGTEALIHAVLHLEQVLTTGGGWQDQVSGLMPGIKVGRS 
               
               
                   
               
               
                 RAQLPLKVEVEEITVPENFVQRKLMAPGCEPLAVHRMMDVLAPYAFGQSL 
               
               
                   
               
               
                 AGAGGGGFLYLLTKEPRQKEVLEAVLAKVEGLGNYSVHLVQVDTQGLSLQ 
               
               
                   
               
               
                 LLGHDAHLCGAGPSEVGNT 
               
               
                   
               
               
                 GenBank Accession No. NP_001100899 (GenBank 
               
               
                   
               
               
                 version dated 05-AUG-2008) 
               
               
                   
               
               
                 mRNA sequence of rat fucose kinase (fucokinase) 
               
               
                 (SEQ ID NO: 28) 
               
               
                 ATGGACCAGCCAAAGGGGGTCAATTGGACGGTCATTATCCTGACATGCCA 
               
               
                   
               
               
                 GTACAAGGACAGTGTCCAGGTCTTTCAGAGAGAGCTGGAGGTAAGGCAG 
               
               
                   
               
               
                 AAGCGGGAGCAGATCCCTGCCGGGACGATGTTACTGGCTGTGGAGGACCC 
               
               
                   
               
               
                 CCAGACCCGAGTAGGCAGTGGAGGAGCTACTCTCAATGCACTGCTGGTGG 
               
               
                   
               
               
                 CTGCTGAGCACCTGAGTGCCCGAGCTGGCTTCACCGTGGTCACGTCAGAT 
               
               
                   
               
               
                 GTCCTGCACTCGGCTTGGATTCTCATCTTGCACATGGGCCGAGACTTCCC 
               
               
                   
               
               
                 CTTTGATGACTGTGGCAGGGCCTTCACTTGCCTCCCTGTGGAGAATCCAC 
               
               
                   
               
               
                 AGGCCCCTGTGGAGGCCTTGGTATGCAACCTGGACTGCCTGTTGGATATC 
               
               
                   
               
               
                 ATGACCCACCGGCTGGGTCCAGGATCCCCACCAGGTGTGTGGGTCTGCAG 
               
               
                   
               
               
                 CACCGACATGCTTCTGTCTGTTCCTCCAAACCCTGGGATCAGTTGGGATG 
               
               
                   
               
               
                 GCTTCCGGGGAACCAGAGTGATCGCCTTTCCTGGGAGCCTGGCCTACGCT 
               
               
                   
               
               
                 CTAAACCACGGGGTCTACCTCACTGACTCGCAGGGCGTGGTTTTGGACAT 
               
               
                   
               
               
                 TTACTACCAGGGCACTAAGGCAGAGATACAACGGTGTGTCAGGCCTGATG 
               
               
                   
               
               
                 GACTGGTACCACTGGTCTCTGGGGTTGTCTTCTTCTCTGTGGAGACTGCT 
               
               
                   
               
               
                 GAGCACCTCCTAGCCACCCACGTGAGCCCACCGCTGGACGCCTGCACCTA 
               
               
                   
               
               
                 TATGGGCTTGGACTCTGGAGCCCAGCCTGTGCAGCTGTCTCTGTTTTTCG 
               
               
                   
               
               
                 ACATCCTGCTCTGCATGGCTCGGAATATGAGCAGGGAGAACTTCGTGGCT 
               
               
                   
               
               
                 GGGCGGCCCCCGGAGATGGGGCAAGGTGACCCGGATGTAGCACGTTACCT 
               
               
                   
               
               
                 GAAGGGAGCCCGGGCCCAGCTGTGGAGGGAGCTTCGAGATCAGCCCCTCA 
               
               
                   
               
               
                 CTATGGTGTATGTCCCTGATGGCGGTTACAGTTACATGACAACTGATGCC 
               
               
                   
               
               
                 ACGGAGTTCCTGCACAGACTCACGATGCCTGGAGTAGCTGTGGCCCAGAT 
               
               
                   
               
               
                 TGTTCACTCTCAGGTGGAGGAGCCACAGCTGCTAGAGGCTACGTGCTCCG 
               
               
                   
               
               
                 TGGTCAGCTGCCTGCTGGAGGGTCCCGTGCACCTGGGGCCTCGAAGTGTC 
               
               
                   
               
               
                 CTGCAGCACTGTCACCTGAGGGGCCCCATTCATATTGGCGCTGGCTGCTT 
               
               
                   
               
               
                 TGTGAGTGGCCTGGATACCGCCCACTCCGAGGCACTGCATGGCCTGGAGC 
               
               
                   
               
               
                 TTCATGACCTCATCCTTCAGGGACACCACATACGGCTGCATGGCTCCCAG 
               
               
                   
               
               
                 AGTCGTGTATTCACTCTTGCTGGCCGTCTGGACAGCTGGGAAAGACAGGG 
               
               
                   
               
               
                 GGCAGGCATGTATCTCAACATGTCCTGGAATGAGTTCTTCAAGAAGACAG 
               
               
                   
               
               
                 GCATTCGAGACTGGGACCTGTGGGACCCAGATACACCCCTCTCAGATCGA 
               
               
                   
               
               
                 TGCCTTCTCAGTGCCCGCCTTTTCCCTGTGCTCCACCCCACGAGGGCTCT 
               
               
                   
               
               
                 GGGGCCCCAGGATGTGCTGTGGATGCTGCATCCTCATAAGGACAGAGGCG 
               
               
                   
               
               
                 AGGCCCTGCGTGCCTGGAGAGCCTCCTGGCGTCTGTCCTGGGAGCAGCTG 
               
               
                   
               
               
                 CAACCTCGCCTGGACCGGGCTGCCACACTGGACTTCCGTCGGGATCTGTT 
               
               
                   
               
               
                 CTTCCGCCAGGCCTTGCAGAAGGCGAGGCATGTGTTAGAGGCCCGGCAGG 
               
               
                   
               
               
                 ACCTCTGCCTACATCCACTGATCCGGGCTGCTGTCGGTGAAGGTTGCTCT 
               
               
                   
               
               
                 GGGCCCCTGCTGGCCACACTTGACAAGGTTGCAGCAGGGGCAGAAGATCC 
               
               
                   
               
               
                 TGGTGTGGCAGCCCGGGCTCTGGCTTGTGTGGCAGATGTACTCGGCTGCA 
               
               
                   
               
               
                 TGGCAGAGGGCCAAGGAGGCTTGCGCAGTGGGCCAGCTGCCAACCCTGAG 
               
               
                   
               
               
                 TGGATTCAGCCTTTCTCATACTTGGAACGTGGAGACCTCATGAGGGGTGT 
               
               
                   
               
               
                 GGAGGCACTTGCCCAGGAAAGAGAGAAGTGGCTGACCAGGCCTGCCTTGT 
               
               
                   
               
               
                 TGGTTCGAGCTGCCCGCCATTATGAGGGGGCTGAGCAGATCCTGATCCGA 
               
               
                   
               
               
                 CAGGCTGTGATGACAGCCCGGCACTTCGTCTCCACCCAGCCAGTGGAATT 
               
               
                   
               
               
                 GCCAGCACCTGGGCAGTGGGTGGTGACTGAGTGCCCAGCCCGTGTGGATT 
               
               
                   
               
               
                 TCTCTGGGGGCTGGAGTGACACACCACCCATTGCCTATGAGCTTGGTGGA 
               
               
                   
               
               
                 GCAGTATTGGGCCTGGCTGTTCGGGTGGATGGCCGCCGGCCCATCGGGGC 
               
               
                   
               
               
                 CAAGGCACGCCGCATCCTAGAGCCTGAGCTCTGGCTGGCAGTGGGACCTC 
               
               
                   
               
               
                 GACAGGATGAGATGACCGTGAAGATAGTGTGCCGGAGCCTTGATGACCTG 
               
               
                   
               
               
                 CAGGATTACTGCCAGCCTCATGCCCCAGGTGCCTTGCTGAAGGCAGCCTT 
               
               
                   
               
               
                 TATCTGTGCGGATATTGTGCATGTCAACTCAGAGGTCCCTCTGCATGAAC 
               
               
                   
               
               
                 AGTTGCTACGCTCGTTTAATGGTGGCTTTGAGCTGCACACATGGTCAGAG 
               
               
                   
               
               
                 CTGCCACACGGCTCTGGTCTTGGCACTAGCAGCATCTTGGCAGGGGCTGC 
               
               
                   
               
               
                 CCTGGCTGCTTTGCAGCGGGCTGCAGGCCGGACAGTGGGCACAGAGGCTC 
               
               
                   
               
               
                 TCATCCATGCAGTGTTGCACCTGGAGCAGGTGCTCACCACAGGAGGTGGC 
               
               
                   
               
               
                 TGGCAGGACCAAGTGAGTGGCCTAATGCCTGGCATCAAGGTGGGGCGCTC 
               
               
                   
               
               
                 TCGGGCACAGCTGCCCCTAAAGGTGGAGGTGGAGGAAATCACTGTGCCTG 
               
               
                   
               
               
                 AGAACTTTGTCCAGAGGAAGCTTATGGCCCCAGGCTGTGAGCCGCTGGCT 
               
               
                   
               
               
                 GTGCATCGGATGATGGATGTCCTGGCCCCTTATGCCTTCGGCCAAAGTCT 
               
               
                   
               
               
                 GGCAGGGGCAGGCGGTGGGGGCTTTCTCTATCTGTTGACCAAGGAACCCC 
               
               
                   
               
               
                 GGCAGAAAGAGGTCCTAGAAGCTGTGCTGGCCAAGGTGGAGGGCCTCGGC 
               
               
                   
               
               
                 AACTACAGCGTCCACCTGGTGCAAGTGGACACTCAGGGCCTGAGCCTGCA 
               
               
                   
               
               
                 GCTGCTAGGACATGACGCCCATCTTTGCGGGGCTGGGCCCTCTGAAGTGG 
               
               
                   
               
               
                 GCAACACCTAG 
               
               
                   
               
               
                 GenBank Accession No. NP_001100899 (GenBank 
               
               
                   
               
               
                 version dated 05-AUG-2008) 
               
             
          
         
       
     
       Fucosyltransferases 
       [0533]    Fucosylated glycans are synthesized by fucosyltransferases, using GDP-fucose as the activated sugar-nucleotide donor. Thirteen fucosyltransferase genes have thus far been identified in the human genome, and include FUT8, FUT4, FUT7, FUT3 and FUT9. FUT8 is an α(1,6)-fucosyltransferase that directs addition of fucose to asparagine-linked GlcNAc moieties, resulting in core fucosylation. 
         [0000]    
       
         
               
               
             
           
               
                 Protein sequence of human fucosyltransferase 8 (α(1,6)-fucosyltransferase) 
                   
               
               
                 (SEQ ID NO: 29) 
                   
               
               
                 MAITVSLVNNKRKIVVLAQPTTVKRKRITPYKSIMTDLYYLSQTDGAGDWRE 
                   
               
               
                   
               
               
                 KEAKDLTELVQRRITYLQNPKDCSKAKKLVCNINKGCGYGCQLHHVVYCFM 
               
               
                   
               
               
                 IAYGTQRTLILESQNWRYATGGWETVFRPVSETCTDRSGISTGHWSGEVKDK 
               
               
                   
               
               
                 NVQVVELPIVDSLHPRPPYLPLAVPEDLADRLVRVHGDPAVWWVSQFVKYLI 
               
               
                   
               
               
                 RPQPWLEKEIEEATKKLGFKHPVIGVHVRRTDKVGTEAAFHPIEEYMVHVEE 
               
               
                   
               
               
                 HFQLLARRMQVDKKRVYLATDDPSLLKEAKTKYPNYEFISDNSISWSAGLHN 
               
               
                   
               
               
                 RYTENSLRGVILDIHFLSQADFLVCTFSSQVCRVAYEIMQTLHPDASANFHSL 
               
               
                   
               
               
                 DDIYYFGGQNAHNQIAIYAHQPRTADEIPMEPGDIIGVAGNHWDGYSKGVNR 
               
               
                   
               
               
                 KLGRTGLYPSYKVREKIETVKYPTYPEAEK 
               
               
                 GenBank Accession No. NP_004480 (GenBank version dated 22 OCT. 2008) 
               
               
                   
               
               
                 mRNA sequence of human fucosyltransferase 8 (α1,6)-fucosyltransferase) 
               
               
                 (SEQ ID NO: 30) 
                   
               
               
                 ggccgacccgagcagccggttccctcctctccaggccccctccccatcccacccccgccgcctggccccagccgaccc 
                   
               
               
                   
               
               
                 gtcccttcgtctccccgcggaatggggccggcactgctcagggtcgcgcgccctggacccagctcgctctcggtctcgcg 
               
               
                   
               
               
                 ctgtcagcgactgcccggctcgcgccgcctcgcgctctgcctcagtcagtggcgccgaaggctccgttaagcggcggcg 
               
               
                   
               
               
                 gcggttcctgtttccgtttcttcctctccgttcggtcgggagtagcatcctccactcagccacccttcccactcccccatcgtgg 
               
               
                   
               
               
                 ggcagctgcggctgagggctgtggctttggcagctgcgacggggagcggcggagaccgcctctgctcccgcctggggt 
               
               
                   
               
               
                 tgctgcttttgctcagaggacatccatgaccctaatggtctttttgttcaagataaagtgattttttgcctttgttgattaactggac 
               
               
                   
               
               
                 aaattcaggataccagaaggccctattgatcaggggccagctataggaagagtacgcgttttagaagagcagcttgttaag 
               
               
                   
               
               
                 gccaaagaacagattgaaaattacaagaaacagaccagaaatggtctggggaaggatcatgaaatcctgaggaggagga 
               
               
                   
               
               
                 ttgaaaatggagctaaagagctctggtttttcctacagagtgaattgaagaaattaaagaacttagaaggaaatgaactccaa 
               
               
                   
               
               
                 agacatgcagatgaatttcttttggatttaggacatcatgaaaggattctgatggcaattactgtctcattagtgaacaataaaa 
               
               
                   
               
               
                 gaaaaattgttgtattagcacaacctactactgtgaagaggaaaagaattaccccatacaagtctataatgacggatctatact 
               
               
                   
               
               
                 acctcagtcagacagatggagcaggtgattggcgggaaaaagaggccaaagatctgacagaactggttcagcggagaat 
               
               
                   
               
               
                 aacatatcttcagaatcccaaggactgcagcaaagccaaaaagctggtgtgtaatatcaacaaaggctgtggctatggctgt 
               
               
                   
               
               
                 cagctccatcatgtggtctactgcttcatgattgcatatggcacccagcgaacactcatcttggaatctcagaattggcgctat 
               
               
                   
               
               
                 gctactggtggatgggagactgtatttaggcctgtaagtgagacatgcacagacagatctggcatctccactggacactggt 
               
               
                   
               
               
                 caggtgaagtgaaggacaaaaatgttcaagtggtcgagcttcccattgtagacagtcttcatccccgtcctccatatttaccct 
               
               
                   
               
               
                 tggctgtaccagaagacctcgcagatcgacttgtacgagtgcatggtgaccctgcagtgtggtgggtgtctcagtttgtcaa 
               
               
                   
               
               
                 atacttgatccgcccacagccttggctagaaaaagaaatagaagaagccaccaagaagcttggcttcaaacatccagttatt 
               
               
                   
               
               
                 ggagtccatgtcagacgcacagacaaagtgggaacagaagctgccttccatcccattgaagagtacatggtgcatgttgaa 
               
               
                   
               
               
                 gaacattttcagcttcttgcacgcagaatgcaagtggacaaaaaaagagtgtatttggccacagatgacccttctttattaaag 
               
               
                   
               
               
                 gaggcaaaaacaaagtaccccaattatgaatttattagtgataactctatttcctggtcagctggactgcacaatcgatacaca 
               
               
                   
               
               
                 gaaaattcacttcgtggagtgatcctggatatacattttctctctcaggcagacttcctagtgtgtactttttcatcccaggtctgt 
               
               
                   
               
               
                 cgagttgcttatgaaattatgcaaacactacatcctgatgcctctgcaaacttccattctttagatgacatctactattttggggg 
               
               
                   
               
               
                 ccagaatgcccacaatcaaattgccatttatgctcaccaaccccgaactgcagatgaaattcccatggaacctggagatatc 
               
               
                   
               
               
                 attggtgtggctggaaatcattgggatggctattctaaaggtgtcaacaggaaattgggaaggacgggcctatatccctccta 
               
               
                   
               
               
                 caaagttcgagagaagatagaaacggtcaagtaccccacatatcctgaggctgagaaataaagctcagatggaagagata 
               
               
                   
               
               
                 aacgaccaaactcagttcgaccaaactcagttcaaaccatttcagccaaactgtagatgaagagggctctgatctaacaaaa 
               
               
                   
               
               
                 taaggttatatgagtagatactctcagcaccaagagcagctgggaactgacataggcttcaattggtggaattcctctttaaca 
               
               
                   
               
               
                 agggctgcaatgccctcatacccatgcacagtacaataatgtactcacatataacatgcaaacaggttgttttctactttgccc 
               
               
                   
               
               
                 ctttcagtatgtccccataagacaaacactgccatattgtgtaatttaagtgacacagacattttgtgtgagacttaaaacatggt 
               
               
                   
               
               
                 gcctatatctgagagacctgtgtgaactattgagaagatcggaacagctccttactctgaggaagttgattcttatttgatggtg 
               
               
                   
               
               
                 gtattgtgaccactgaattcactccagtcaacagattcagaatgagaatggacgtttggtttttttttgtttttgtttttgttttttccttt 
               
               
                   
               
               
                 ataaggttgtctgtttttttttttttaaataattgcatcagttcattgacctcatcattaataagtgaagaatacatcagaaaataaaat 
               
               
                   
               
               
                 attcactctccattagaaaattttgtaaaacaatgccatgaacaaattctttagtactcaatgtttctggacattctctttgataaca 
               
               
                   
               
               
                 aaaaataaattttaaaaaggaattttgtaaagtttctagaattttatatcattggatgatatgttgatcagccttatgtggaagaact 
               
               
                   
               
               
                 gtgataaaaagaggagctttttagtttttcagcttaaaaaaa 
               
               
                 GenBank Accession No. NP_004480 (GenBank version dated 22 OCT. 2008) 
               
               
                   
               
               
                 Protein sequence of rat fucosyltransferase 8 (α1,6)-fucosyltransferase) 
               
               
                 (SEQ ID NO: 31) 
                   
               
               
                 MRAWTGSWRWIMLILFAWGTLLFYIGGHLVRDNDHPDHSSRELSKILAKLER 
                   
               
               
                   
               
               
                 LKQQNEDLRRMAESLR1PEGPIDQGTATGRVRVLEEQLVKAKEQIENYKKQA 
               
               
                   
               
               
                 RNGLGKDHELLRRRIENGAKELWFFLQSELKKLKHLEGNELQRHADEILLDL 
               
               
                   
               
               
                 GHHERSIMTDLYYLSQTDGAGDWREKEAKDLTELVQRRITYLQNPKDCSKA 
               
               
                   
               
               
                 RKLVCNINKGCGYGCQLHHVVYCFMIAYGTQRTLILESQNWRYATGGWETV 
               
               
                   
               
               
                 FRPVSETCTDRSGLSTGHWSGEVNDKNIQVVELPIVDSLHPRPPYLPLAVPEDL 
               
               
                   
               
               
                 ADRLVRVHGDPAVWWVSQFVKYLIRPQPWLEKEIEEATKKLGFKHPVIGVH 
               
               
                   
               
               
                 VRRTDKVGTEAAFHPIEEYMVHVEEHFQLLARRMQVDKKRVYLATDDPALL 
               
               
                   
               
               
                 KEAKTKYSNYEFISDNSISWSAGLHNRYTENSLRGVILDIHFLSQADFLVCTFS 
               
               
                   
               
               
                 SQVCRVAYEIMQTLHPDASANFHSLDDIYYFGGQNAHNQIAVYPHKPRTDEEI 
               
               
                   
               
               
                 PMEPGDIIGVAGNHWDGYSKGVNRKLGKTGLYPSYKVREKIETVKYPTYPEA 
               
               
                   
               
               
                 EK 
               
               
                 GenBank Accession No. NP_001002289 (GenBank version dated 5 OCT. 2008) 
               
               
                   
               
               
                 mRNA sequence of rat fucosyltransferase 8 (α1,6)-fucosyltransferase) 
               
               
                 (SEQ ID NO: 32) 
                   
               
               
                 atgcgggcatggactggttcctggcgttggattatgctcattctttttgcctgggggaccttgttgttttatataggtggtcatttg 
                   
               
               
                   
               
               
                 gttcgagataatgaccaccctgatcactctagcagagaactctccaagattcttgcaaagcttgaacgcttaaaacaacaaaa 
               
               
                   
               
               
                 tgaagacttgaggcgaatggctgagtctctacgaataccagaaggccccattgaccaggggacggctacgggaagagtc 
               
               
                   
               
               
                 cgtgttttagaagaacagcttgttaaggccaaagaacagattgaaaattacaagaaacaagccagaaatggtctggggaag 
               
               
                   
               
               
                 gatcatgaactcttaaggaggaggattgaaaatggagctaaagagctctggttttttctacaaagtgaactgaagaaattaaa 
               
               
                   
               
               
                 gcatctagaaggaaatgaactccaaagacatgcagatgaaattcttttggatttaggacaccatgaaaggtctatcatgacgg 
               
               
                   
               
               
                 atctatactacctcagtcaaacagatggagcaggggattggcgtgaaaaagaggccaaagatctgacagagctggtccag 
               
               
                   
               
               
                 cggagaataacttatctccagaatcccaaggactgcagcaaagccaggaagctggtgtgtaacatcaataagggctgtgg 
               
               
                   
               
               
                 ctatggttgccaactccatcacgtggtctactgtttcatgattgcttatggcacccagcgaacactcatcttggaatctcagaatt 
               
               
                   
               
               
                 ggcgctatgctactggtggatgggagactgtgtttagacctgtaagtgagacatgcacagacagatctggcctctccactgg 
               
               
                   
               
               
                 acactggtcaggtgaagtgaatgacaaaaatattcaagtggtggagctccccattgtagacagcctccatcctcggcctcctt 
               
               
                   
               
               
                 acttaccactggctgttccagaagaccttgcagatcgactcgtaagagtccatggtgatcctgcagtgtggtgggtgtccca 
               
               
                   
               
               
                 gttcgtcaaatatttgattcgtccacaaccttggctagaaaaggaaatagaagaagccaccaagaagcttggcttcaaacatc 
               
               
                   
               
               
                 cagtcattggagtccatgtcagacgcacagacaaagtgggaacagaggcagccttccatcccatcgaagagtacatggta 
               
               
                   
               
               
                 catgttgaagaacattttcagcttctcgcacgcagaatgcaagtggataaaaaaagagtatatctggctaccgatgaccctgc 
               
               
                   
               
               
                 tttgttaaaggaggcaaagacaaagtactccaattatgaatttattagtgataactctatttcttggtcagctggattacacaatc 
               
               
                   
               
               
                 ggtacacagaaaattcacttcggggcgtgatcctggatatacactttctctctcaggctgacttcctagtgtgtactttttcatcc 
               
               
                   
               
               
                 caggtctgtcgggttgcttatgaaatcatgcaaaccctgcatcctgatgcctctgcaaacttccactctttagatgacatctact 
               
               
                   
               
               
                 attttggaggccaaaatgcccacaaccagattgccgtttatcctcacaaacctcgaactgatgaggaaattccaatggaacct 
               
               
                   
               
               
                 ggagatatcattggtgtggctggaaaccattgggatggttattctaaaggtgtcaacagaaaacttggaaaaacaggcttata 
               
               
                   
               
               
                 tccctcctacaaagtccgagagaagatagaaacagtcaagtatcccacatatcctgaagctgaaaaatag 
               
               
                 GenBank Accession No. NM_001002289 (GenBank version dated 5 OCT. 2008) 
               
               
                   
               
               
                 Protein sequence of mouse fucosyltransferase 8 (α1,6)-fucosyltransferase) 
               
               
                 (SEQ ID NO: 33) 
                   
               
               
                 MRAWTGSWRWIMLILFAWGTLLFYIGGHLVRDNDHPDHSSRELSKILAKLER 
                   
               
               
                   
               
               
                 LKQQNEDLRRMAESLR1PEGPIDQGTATGRVRVLEEQLVKAKEQIENYKKQA 
               
               
                   
               
               
                 RNGLGKDHEILRRRIENGAKELWFFLQSELKKLKHLEGNELQRHADEILLDLG 
               
               
                   
               
               
                 HHERSIMTDLYYLSQTDGAGDWREKEAKDLTELVQRRITYLQNPKDCSKAR 
               
               
                   
               
               
                 KLVCNINKGCGYGCQLHHVVYCFMIAYGTQRTLILESQNWRYATGGWETVF 
               
               
                   
               
               
                 RPVSETCTDRSGLSTGHWSGEVNDKNIQVVELPIVDSLHPRPPYLPLAVPEDL 
               
               
                   
               
               
                 ADRLLRVHGDPAVWWVSQFVKYLIRPQPWLEKEIEEATKKLGFKHPVIGVHV 
               
               
                   
               
               
                 RRTDKVGTEAAFHPIEEYMVHVEEHFQLLARRMQVDKKRVYLATDDPTLLK 
               
               
                   
               
               
                 EAKTKYSNYEFISDNSISWSAGLHNRYTENSLRGVILDIHFLSQADFLVCTFSS 
               
               
                   
               
               
                 QVCRVAYEIMQTLHPDASANFHSLDDIYYFGGQNAHNQIAVYPHKPRTEEEIP 
               
               
                   
               
               
                 MEPGDIIGVAGNHWDGYSKGINRKLGKTGLYPSYKVREKIETVKYPTYPEAE 
               
               
                   
               
               
                 K 
               
               
                 GenBank Accession No. NP_058589 (GenBank version dated 04 JAN. 2009) 
               
               
                   
               
               
                 mRNA sequence of mouse fucosyltransferase 8 (A1,6)-fucosyltransferase) 
               
               
                 (SEQ ID NO: 34) 
                   
               
               
                 atgcgggcatggactggttcctggcgttggattatgctcattctttttgcctgggggaccttgttattttatataggtggtcatttg 
                   
               
               
                   
               
               
                 gttcgagataatgaccaccctgatcactccagcagagaactctccaagattcttgcaaagcttgaacgcttaaaacagcaaa 
               
               
                   
               
               
                 atgaagacttgaggcgaatggctgagtctctccgaataccagaaggccccattgaccaggggacagctacaggaagagt 
               
               
                   
               
               
                 ccgtgttttagaagaacagcttgttaaggccaaagaacagattgaaaattacaagaaacaagctagaaatggtctggggaa 
               
               
                   
               
               
                 ggatcatgaaatcttaagaaggaggattgaaaatggagctaaagagctctggttttttctacaaagcgaactgaagaaattaa 
               
               
                   
               
               
                 agcatttagaaggaaatgaactccaaagacatgcagatgaaattcttttggatttaggacaccatgaaaggtctatcatgaca 
               
               
                   
               
               
                 gatctatactacctcagtcaaacagatggagcaggggattggcgtgaaaaagaggccaaagatctgacagagctggtcca 
               
               
                   
               
               
                 gcggagaataacatatctccagaatcctaaggactgcagcaaagccaggaagctggtgtgtaacatcaataaaggctgtg 
               
               
                   
               
               
                 gctatggttgtcaactccatcacgtggtctactgtttcatgattgcttatggcacccagcgaacactcatcttggaatctcagaa 
               
               
                   
               
               
                 ttggcgctatgctactggtggatgggagactgtgtttagacctgtaagtgagacatgtacagacagatctggcctctccactg 
               
               
                   
               
               
                 gacactggtcaggtgaagtaaatgacaaaaacattcaagtggtcgagctccccattgtagacagcctccatcctcggcctcc 
               
               
                   
               
               
                 ttacttaccactggctgttccagaagaccttgcagaccgactcctaagagtccatggtgaccctgcagtgtggtgggtgtccc 
               
               
                   
               
               
                 agtttgtcaaatacttgattcgtccacaaccttggctggaaaaggaaatagaagaagccaccaagaagcttggcttcaaaca 
               
               
                   
               
               
                 tccagttattggagtccatgtcagacgcacagacaaagtgggaacagaagcagccttccaccccatcgaggagtacatgg 
               
               
                   
               
               
                 tacacgttgaagaacattttcagcttctcgcacgcagaatgcaagtggataaaaaaagagtatatctggctactgatgatccta 
               
               
                   
               
               
                 ctttgttaaaggaggcaaagacaaagtactccaattatgaatttattagtgataactctatttcttggtcagctggactacacaat 
               
               
                   
               
               
                 cggtacacagaaaattcacttcggggtgtgatcctggatatacactttctctcacaggctgactttctagtgtgtactttttcatcc 
               
               
                   
               
               
                 caggtctgtcgggttgcttatgaaatcatgcaaaccctgcatcctgatgcctctgcgaacttccattctttggatgacatctact 
               
               
                   
               
               
                 attttggaggccaaaatgcccacaatcagattgctgtttatcctcacaaacctcgaactgaagaggaaattccaatggaacct 
               
               
                   
               
               
                 ggagatatcattggtgtggctggaaaccattgggatggttattctaaaggtatcaacagaaaacttggaaaaacaggcttata 
               
               
                   
               
               
                 tccctcctacaaagtccgagagaagatagaaacagtcaagtatcccacatatcctgaagctgaaaaatag 
               
               
                 GenBank Accession No. NM_016893 (GenBank version dated 04 JAN. 2009) 
               
             
          
         
       
     
       GDP-Fucose Transporters 
       [0534]    Fucosylated glycans are synthesized by fucosyltransferases in the Golgi apparatus, while GDP-fucose is synthesized in the cytosol. Thus, GDP-fucose must be translocated to the Golgi by a GDP-fucose transporter, such as GDP-fucose transporter 1 (FUCT1). 
         [0000]    
       
         
               
             
           
               
                 Protein sequence of human GDP-fucose transporter 1 
               
               
                 (FUCT1) 
               
               
                 (SEQ ID NO: 35) 
               
               
                 MNRAPLKRSRILHMALTGASDPSAEAEANGEKPFLLRALQIALVVSLYW 
               
               
                   
               
               
                 VTSISMVFLNKYLLDSPSLRLDTPIFVTFYQCLVTTLLCKGLSALAACC 
               
               
                   
               
               
                 PGAVDFPSLRLDLRVARSVLPLSVVFIGMITFNNLCLKYVGVAFYNVGR 
               
               
                   
               
               
                 SLTTVFNVLLSYLLLKQTTSFYALLTCGIIIGGFWLGVDQEGAEGTLSW 
               
               
                   
               
               
                 LGTVFGVLASLCVSLNAIYTTKVLPAVDGSIWRLTFYNNVNACILFLPL 
               
               
                   
               
               
                 LLLLGELQALRDFAQLGSAHFWGMMTLGGLFGFAIGYVTGLQIKFTSPL 
               
               
                   
               
               
                 THNVSGTAKACAQTVLAVLYYEETKSFLWWTSNMMVLGGSSAYTWVRGW 
               
               
                   
               
               
                 EMKKTPEEPSPKDSEKSAMGV 
               
               
                 GenBank Accession No. NP_060859 (GenBank version 
               
               
                 dated 27 FEB. 2009) 
               
               
                   
               
               
                 mRNA sequence of human GDP-fucose transporter 1 
               
               
                 (FUCT1) 
               
               
                 (SEQ ID NO: 36) 
               
               
                 ATGAATAGGGCCCCTCTGAAGCGGTCCAGGATCCTGCACATGGCGCTGA 
               
               
                   
               
               
                 CCGGGGCCTCAGACCCCTCTGCAGAGGCAGAGGCCAACGGGGAGAAGCC 
               
               
                   
               
               
                 CTTTCTGCTGCGGGCATTGCAGATCGCGCTGGTGGTCTCCCTCTACTGG 
               
               
                   
               
               
                 GTCACCTCCATCTCCATGGTGTTCCTTAATAAGTACCTGCTGGACAGCC 
               
               
                   
               
               
                 CCTCCCTGCGGCTGGACACCCCCATCTTCGTCACCTTCTACCAGTGCCT 
               
               
                   
               
               
                 GGTGACCACGCTGCTGTGCAAAGGCCTCAGCGCTCTGGCCGCCTGCTGC 
               
               
                   
               
               
                 CCTGGTGCCGTGGACTTCCCCAGCTTGCGCCTGGACCTCAGGGTGGCCC 
               
               
                   
               
               
                 GCAGCGTCCTGCCCCTGTCGGTGGTCTTCATCGGCATGATCACCTTCAA 
               
               
                   
               
               
                 TAACCTCTGCCTCAAGTACGTCGGTGTGGCCTTCTACAATGTGGGCCGC 
               
               
                   
               
               
                 TCACTCACCACCGTCTTCAACGTGCTGCTCTCCTACCTGCTGCTCAAGC 
               
               
                   
               
               
                 AGACCACCTCCTTCTATGCCCTGCTCACCTGCGGTATCATCATCGGGGG 
               
               
                   
               
               
                 CTTCTGGCTTGGTGTGGACCAGGAGGGGGCAGAAGGCACCCTGTCGTGG 
               
               
                   
               
               
                 CTGGGCACCGTCTTCGGCGTGCTGGCTAGCCTCTGTGTCTCGCTCAACG 
               
               
                   
               
               
                 CCATCTACACCACGAAGGTGCTCCCGGCGGTGGACGGCAGCATCTGGCG 
               
               
                   
               
               
                 CCTGACTTTCTACAACAACGTCAACGCCTGCATCCTCTTCCTGCCCCTG 
               
               
                   
               
               
                 CTCCTGCTGCTCGGGGAGCTTCAGGCCCTGCGTGACTTTGCCCAGCTGG 
               
               
                   
               
               
                 GCAGTGCCCACTTCTGGGGGATGATGACGCTGGGCGGCCTGTTTGGCTT 
               
               
                   
               
               
                 TGCCATCGGCTACGTGACAGGACTGCAGATCAAGTTCACCAGTCCGCTG 
               
               
                   
               
               
                 ACCCACAATGTGTCGGGCACGGCCAAGGCCTGTGCCCAGACAGTGCTGG 
               
               
                   
               
               
                 CCGTGCTCTACTACGAGGAGACCAAGAGCTTCCTCTGGTGGACGAGCAA 
               
               
                   
               
               
                 CATGATGGTGCTGGGCGGCTCCTCCGCCTACACCTGGGTCAGGGGCTGG 
               
               
                   
               
               
                 GAGATGAAGAAGACTCCGGAGGAGCCCAGCCCCAAAGACAGCGAGAAGA 
               
               
                   
               
               
                 GCGCCATGGGGGTGTGA 
               
               
                 GenBank Accession No. NM_018389 (GenBank version 
               
               
                 dated 27 FEB. 2009) 
               
               
                   
               
               
                 Protein sequence of mouse GDP-fucose transporter 1 
               
               
                 (FUCT1) 
               
               
                 (SEQ ID NO: 37) 
               
               
                 MNRAPLKRSRILRMALTGVSAVSEESESGNKPFLLRALQIALVVSSLYW 
               
               
                   
               
               
                 VTSISMVFLNKYLLDSPSLQLDTPIFVTFYQCLVTSLLCKGLSTLATCC 
               
               
                   
               
               
                 PGMVDFPTLNLDLKVARSVLPLSVVFIGMITFNNLCLKYVGVPFYNVGR 
               
               
                   
               
               
                 SLTTVFNVLLSYLLLKQTTSFYALLTCGVIIGGFWLGIDQEGAEGTLSL 
               
               
                   
               
               
                 TGTIFGVLASLCVSLNAIYTKKVLPAVDHSIWRLTFYNNVNACVLFLPL 
               
               
                   
               
               
                 MIVLGELRALLAFTHLSSAHFWLMMTLGGLFGFAIGYVTGLQIKFTSPL 
               
               
                   
               
               
                 THNVSGTAKACAQTVLAVLYYEEIKSFLWWTSNLMVLGGSSAYTWVRGW 
               
               
                   
               
               
                 EMQKTQEDPSSKDGEKSAIRV 
               
               
                 GenBank Accession No. NP_997597 (GenBank version 
               
               
                 dated 21-SEP-2008) 
               
               
                   
               
               
                 mRNA sequence of mouse GDP-fucose transporter 1 
               
               
                 (FUCT1) 
               
               
                 (SEQ ID NO: 38) 
               
               
                 ATGAACAGGGCGCCTCTGAAGCGGTCCAGGATCCTGCGCATGGCGCTGA 
               
               
                   
               
               
                 CTGGAGTCTCTGCTGTCTCCGAGGAGTCAGAGAGCGGGAACAAGCCATT 
               
               
                   
               
               
                 TCTGCTCCGGGCTCTGCAGATCGCGCTGGTGGTCTCTCTCTACTGGGTC 
               
               
                   
               
               
                 ACCTCCATTTCCATGGTATTCCTCAACAAGTACCTGCTGGACAGCCCCT 
               
               
                   
               
               
                 CCCTGCAGCTGGATACCCCCATTTTTGTCACCTTCTACCAATGCCTGGT 
               
               
                   
               
               
                 GACCTCACTGCTGTGCAAGGGCCTCAGCACTCTGGCCACCTGCTGCCCC 
               
               
                   
               
               
                 GGCATGGTAGACTTCCCCACCCTAAACCTGGACCTCAAGGTGGCCCGAA 
               
               
                   
               
               
                 GTGTGCTGCCGCTGTCAGTGGTCTTTATCGGCATGATAACCTTCAATAA 
               
               
                   
               
               
                 CCTCTGCCTCAAGTACGTAGGGGTGCCCTTCTACAACGTGGGACGCTCG 
               
               
                   
               
               
                 CTCACCACCGTGTTCAACGTTCTTCTCTCCTACCTGCTGCTCAAACAGA 
               
               
                   
               
               
                 CCACTTCCTTCTATGCCCTGCTCACCTGCGGCGTCATCATTGGTGGTTT 
               
               
                   
               
               
                 CTGGCTGGGTATAGACCAAGAAGGAGCTGAGGGAACCTTGTCCCTGACG 
               
               
                   
               
               
                 GGCACCATCTTCGGGGTGCTGGCCAGCCTCTGCGTCTCCCTCAATGCCA 
               
               
                   
               
               
                 TCTATACCAAGAAGGTGCTCCCTGCAGTAGACCACAGTATCTGGCGCCT 
               
               
                   
               
               
                 AACCTTCTATAACAATGTCAATGCCTGCGTGCTCTTCTTGCCCCTGATG 
               
               
                   
               
               
                 ATAGTGCTGGGCGAGCTCCGTGCCCTCCTGGCCTTCACTCATCTGAGCA 
               
               
                   
               
               
                 GTGCCCACTTCTGGCTCATGATGACGCTGGGTGGCCTGTTTGGCTTTGC 
               
               
                   
               
               
                 CATCGGCTATGTGACAGGACTGCAGATCAAATTCACCAGTCCCCTGACC 
               
               
                   
               
               
                 CATAACGTGTCAGGCACGGCCAAGGCCTGTGCACAGACAGTGCTGGCCG 
               
               
                   
               
               
                 TGCTCTACTACGAAGAGATTAAGAGCTTCCTGTGGTGGACAAGCAACCT 
               
               
                   
               
               
                 GATGGTGCTGGGTGGCTCCTCCGCCTACACCTGGGTCAGGGGCTGGGAG 
               
               
                   
               
               
                 ATGCAGAAGACCCAGGAGGACCCCAGCTCCAAAGATGGTGAGAAGAGTG 
               
               
                   
               
               
                 CTATCAGGGTGTGA 
               
               
                 GenBank Accession No. NM_211358 (GenBank version 
               
               
                 dated 21 SEP. 2008) 
               
               
                   
               
               
                 Protein sequence of rat GDP-fucose transporter 1 
               
               
                 (FUCT1) 
               
               
                 (SEQ ID NO: 39) 
               
               
                 MNRVPLKRSRILRMALTGASAVSEEADSENKPFLLRALQIALVVSLYWV 
               
               
                   
               
               
                 TSISMVFLNKYLLDSPSLQLDTPIFVTFYQCLVTSLLCKGLSTLATCCP 
               
               
                   
               
               
                 GMVDFPTLNLDLKVARSVLPLSVVFIGMITFNNLCLKYVGVAFYNVGRS 
               
               
                   
               
               
                 LTTVFNVLLSYLLLKQTTSFYALLTCAIIIGGFWLGIDQEGAEGTLSLT 
               
               
                   
               
               
                 GTIFGVLASLCVSLNAIYTKKVLPAVDHSIWRLTFYNNVNACVLFLPLM 
               
               
                   
               
               
                 VVLGELHALLAFAHLNSAHFWVMMTLGGLFGFAIGYVTGLQIKFTSPLT 
               
               
                   
               
               
                 HNVSGTAKACAQTVLAVLYYEEIKSFLWWTSNLMVLGGSSAYTWVRGWE 
               
               
                   
               
               
                 MQKTQEDPSSKEGEKSAIGV 
               
               
                 GenBank Accession No. NP_001101218 (GenBank 
               
               
                 version dated 18 FEB. 2009) 
               
               
                   
               
               
                 mRNA sequence of rat GDP-fucose transporter 1 
               
               
                 (FUCT1) 
               
               
                 (SEQ ID NO: 40) 
               
               
                 ATGAACAGGGTCCCTCTGAAGCGGTCCAGGATCCTGCGCATGGCGCTGA 
               
               
                   
               
               
                 CTGGAGCCTCTGCTGTCTCTGAGGAGGCAGACAGCGAGAACAAGCCATT 
               
               
                   
               
               
                 TCTGCTACGGGCTCTGCAGATCGCGCTGGTGGTTTCTCTCTACTGGGTC 
               
               
                   
               
               
                 ACCTCCATCTCCATGGTATTCCTCAACAAGTACCTGCTGGACAGCCCCT 
               
               
                   
               
               
                 CCCTGCAGCTGGATACCCCCATCTTCGTCACCTTCTACCAATGCCTGGT 
               
               
                   
               
               
                 GACCTCACTGCTGTGCAAGGGCCTCAGCACTCTGGCCACCTGCTGCCCT 
               
               
                   
               
               
                 GGCATGGTAGACTTCCCCACCCTAAACCTGGACCTCAAGGTGGCCCGAA 
               
               
                   
               
               
                 GTGTGCTGCCGCTGTCCGTGGTCTTTATCGGCATGATAACCTTCAATAA 
               
               
                   
               
               
                 CCTCTGCCTCAAGTACGTGGGGGTGGCCTTCTACAACGTGGGACGCTCG 
               
               
                   
               
               
                 CTCACTACCGTGTTCAATGTGCTTCTCTCCTACCTGCTGCTTAAACAGA 
               
               
                   
               
               
                 CCACTTCCTTTTATGCCCTGCTCACCTGTGCCATCATCATTGGTGGTTT 
               
               
                   
               
               
                 CTGGCTGGGAATAGATCAAGAGGGAGCTGAGGGCACCCTGTCCCTGACG 
               
               
                   
               
               
                 GGCACCATCTTCGGGGTGCTGGCCAGCCTCTGTGTCTCACTCAATGCCA 
               
               
                   
               
               
                 TCTACACCAAGAAGGTGCTCCCTGCCGTAGACCACAGTATCTGGCGCCT 
               
               
                   
               
               
                 AACCTTCTATAACAACGTCAACGCCTGTGTGCTCTTCTTGCCCCTGATG 
               
               
                   
               
               
                 GTAGTGCTGGGCGAGCTCCATGCTCTCCTGGCCTTCGCTCATCTGAACA 
               
               
                   
               
               
                 GCGCCCACTTCTGGGTCATGATGACGCTGGGTGGACTCTTCGGCTTTGC 
               
               
                   
               
               
                 CATTGGCTATGTGACAGGACTGCAGATCAAATTCACCAGTCCCCTGACC 
               
               
                   
               
               
                 CATAATGTGTCGGGCACAGCCAAGGCCTGTGCACAGACAGTGCTGGCTG 
               
               
                   
               
               
                 TGCTCTACTATGAAGAGATTAAGAGCTTCCTGTGGTGGACAAGCAACTT 
               
               
                   
               
               
                 GATGGTGCTGGGTGGCTCCTCTGCCTACACCTGGGTCAGGGGCTGGGAG 
               
               
                   
               
               
                 ATGCAGAAGACCCAGGAGGACCCCAGCTCCAAAGAGGGTGAGAAGAGTG 
               
               
                   
               
               
                 CTATCGGGGTGTGA 
               
               
                 GenBank Accession No. NM_001107748 (GenBank 
               
               
                 version dated 18 FEB. 2009) 
               
               
                   
               
               
                 Protein sequence of Chinese hamster GDP-fucose 
               
               
                 transporter 1 (FUCT1) 
               
               
                 (SEQ ID NO: 41) 
               
               
                 MNRAPLKRSRILRMALTGGSTASEEADEDSRNKPFLLRALQIALVVSLY 
               
               
                   
               
               
                 WVTSISMVFLNKYLLDSPSLQLDTPIFVTFYQCLVTSLLCKGLSTLATC 
               
               
                   
               
               
                 CPGTVDFPTLNLDLKVARSVLPLSVVFIGMISFNNLCLKYVGVAFYNVG 
               
               
                   
               
               
                 RSLTTVFNVLLSYLLLKQTTSFYALLTCGIIIGGFWLGIDQEGAEGTLS 
               
               
                   
               
               
                 LIGTIFGVLASLCVSLNAIYTKKVLPAVDNSIWRLTFYNNVNACVLFLP 
               
               
                   
               
               
                 LMVLLGELRALLDFAHLYSAHFWLMMTLGGLFGFAIGYVTGLQIKFTSP 
               
               
                   
               
               
                 LTHNVSGTAKACAQTVLAVLYYEETKSFLWWTSNLMVLGGSSAYTWVRG 
               
               
                   
               
               
                 WEMQKTQEDPSSKEGEKSAIRV 
               
               
                 GenBank Accession No. BAE16173 (GenBank version 
               
               
                 dated 12 SEP. 2008) 
               
               
                   
               
               
                 mRNA sequence of Chinese hamster GDP-fucose 
               
               
                 transporter 1 (FUCT1) 
               
               
                 (SEQ ID NO: 42) 
               
               
                 ATGAACAGGGCGCCTCTGAAGCGGTCCAGGATCCTGCGCATGGCGCTGA 
               
               
                   
               
               
                 CTGGAGGCTCCACTGCCTCTGAGGAGGCAGATGAGGACAGCAGGAACAA 
               
               
                   
               
               
                 GCCGTTTCTGCTGCGGGCGCTGCAGATCGCGCTGGTCGTCTCTCTCTAC 
               
               
                   
               
               
                 TGGGTCACCTCCATCTCCATGGTATTCCTCAACAAGTACCTGCTGGACA 
               
               
                   
               
               
                 GCCCCTCCCTGCAGCTGGATACCCCTATCTTCGTCACTTTCTACCAATG 
               
               
                   
               
               
                 CCTGGTGACCTCTCTGCTGTGCAAGGGCCTCAGCACTCTGGCCACCTGC 
               
               
                   
               
               
                 TGCCCTGGCACCGTTGACTTCCCCACCCTGAACCTGGACCTTAAGGTGG 
               
               
                   
               
               
                 CCCGCAGCGTGCTGCCACTGTCGGTAGTCTTCATTGGCATGATAAGTTT 
               
               
                   
               
               
                 CAATAACCTCTGCCTCAAGTACGTAGGGGTGGCCTTCTACAACGTGGGG 
               
               
                   
               
               
                 CGCTCGCTCACCACCGTGTTCAATGTGCTTCTGTCCTACCTGCTGCTCA 
               
               
                   
               
               
                 AACAGACCACTTCCTTCTATGCCCTGCTCACATGTGGCATCATCATTGG 
               
               
                   
               
               
                 TGGTTTCTGGCTGGGTATAGACCAAGAGGGAGCTGAGGGCACCCTGTCC 
               
               
                   
               
               
                 CTCATAGGCACCATCTTCGGGGTGCTGGCCAGCCTCTGCGTCTCCCTCA 
               
               
                   
               
               
                 ATGCCATCTATACCAAGAAGGTGCTCCCAGCAGTGGACAACAGCATCTG 
               
               
                   
               
               
                 GCGCCTAACCTTCTATAACAATGTCAATGCCTGTGTGCTCTTCTTGCCC 
               
               
                   
               
               
                 CTGATGGTTCTGCTGGGTGAGCTCCGTGCCCTCCTTGACTTTGCTCATC 
               
               
                   
               
               
                 TGTACAGTGCCCACTTCTGGCTCATGATGACGCTGGGTGGCCTCTTCGG 
               
               
                   
               
               
                 CTTTGCCATTGGCTATGTGACAGGACTGCAGATCAAATTCACCAGTCCC 
               
               
                   
               
               
                 CTGACCCACAATGTATCAGGCACAGCCAAGGCCTGTGCGCAGACAGTGC 
               
               
                   
               
               
                 TGGCCGTGCTCTACTATGAAGAGACTAAGAGCTTCCTGTGGTGGACAAG 
               
               
                   
               
               
                 CAACCTGATGGTGCTGGGTGGCTCCTCAGCCTATACCTGGGTCAGGGGC 
               
               
                   
               
               
                 TGGGAGATGCAGAAGACCCAAGAGGACCCCAGCTCCAAAGAGGGTGAGA 
               
               
                   
               
               
                 AGAGTGCTATCAGGGTGTGA 
               
               
                 GenBank Accession No. AB222037 (GenBank version 
               
               
                 dated 12 SEP. 2008) 
               
             
          
         
       
     
         [0535]    Proteins or nucleic acids used in the methods and cells described herein (e.g., GMD, FX, GFPP, fucose kinase, GDP-fucose synthetase, a fucosyltransferase or a GDP-fucose transporter) include mammalian (e.g., human, mouse, rat or hamster) proteins. A protein, nucleic acid or cell can be a primate (e.g., human) protein, nucleic acid or cell. In other embodiments the protein, nucleic acid or cell is a rodent (e.g., a mouse, rat or hamster) protein, nucleic acid or cell. 
         [0536]    A protein sequence, e.g., a protein encoding sequence, can be used to decrease the protein expression in a cell. For example, a decrease in protein expression can be achieved by inactivating the endogenous gene, e.g., in the control or structural regions. A cloned sequence can be used to make a construct that will insert a deletion or other event into an endogenous gene to decrease levels of the protein it expresses. 
         [0537]    The expression of endogenous protein can be decreased by the use of a genetic construct from the same species as the endogenous protein, or from a different species. For example, the expression of an endogenous protein in a mouse cell can be modulated with a construct made from mouse protein or with one made from a protein sequence from another species, e.g., a different rodent species. The protein of a rodent, e.g., a hamster, such as a Chinese hamster, can be manipulated with an allogeneic sequence (from the same species) or a xenogeneic sequence (from a different species). For example, a CHO cell can be manipulated with a Chinese hamster, mouse or rat sequence. 
         [0538]    A nucleic acid sequence from one of the proteins disclosed herein can be used to isolate a gene from a different species. For example, a mouse or rat sequence described herein can be used to make primers to isolate a sequence from another rodent, e.g., a hamster, e.g., a Chinese hamster. That sequence can them be used to modify protein expression in a cell, e.g., in a Chinese hamster cell, such as a CHO cell. 
       Manipulations 
       [0539]    As described above, a manipulation, as used herein, refers to a property of a cell. Examples of manipulations include the presence in or on the cell of an exogenous inhibitor of an enzyme involved in the biosynthesis of GDP-fucose, or a nucleic acid antagonist (e.g., an siRNA) 
         [0540]    A manipulated cell can be, e.g., a vertebrate, mammalian or rodent cell. Primers or other nucleic acids used, e.g., to form or make manipulations, can be, e.g., vertebrate, mammalian or rodent sequences. For example, a rodent primer or other nucleic acid, e.g., a nucleic acid encoding an active or inactivate rodent GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, a fucosyltransferase or GDP-fucose transporter protein, can be used to manipulate a rodent cell. Similarly, a mammalian cell having a manipulation can be made with mammalian nucleic acids, e.g., mammalian primers or a nucleic acid encoding a mammalian GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, a fucosyltransferase or GDP-fucose transporter protein. A sequence from a first species can be used to manipulate a cell of a second species. E.g., a primer or nucleic acid from a first species, e.g., a first rodent species, e.g., a mouse or rat, can be used to manipulate a cell from a second species, e.g., a second rodent species, e.g., a hamster cell, e.g., a CHO cell. 
       Nucleic Acid Antagonists 
       [0541]    In some embodiments, nucleic acid antagonists are used to decrease expression of a target protein, e.g., a protein involved in regulating GDP-fucose levels, e.g., a protein involved in GDP-fucose biosynthesis, a fucosyltransferase or a GDP-fucose transporter. In one embodiment, the nucleic acid antagonist is an siRNA that targets mRNA encoding the target protein. Other types of antagonistic nucleic acids can also be used, e.g., a nucleic acid aptamer, a dsRNA, a ribozyme, a triple-helix former, or an antisense nucleic acid. 
         [0542]    siRNAs can be used to inhibit expression of a protein involved in GDP-fucose biosynthesis, a fucosyltransferase or a GDP-fucose transporter. siRNAs are small double stranded RNAs (dsRNAs) that optionally include overhangs. For example, the duplex region of an siRNA is about 18 to 25 nucleotides in length, e.g., about 19, 20, 21, 22, 23, or 24 nucleotides in length. Typically the siRNA sequences are exactly complementary to the target mRNA. dsRNAs and siRNAs in particular can be used to silence gene expression in mammalian cells (e.g., human cells). See, e.g., Clemens, J. C. et al. (2000)  Proc. Natl. Sci. USA  97, 6499-6503; Billy, E. et al. (2001)  Proc. Natl. Sci. USA  98, 14428-14433; Elbashir et al. (2001)  Nature  411(6836):494-8; Yang, D. et al. (2002)  Proc. Natl. Acad. Sci. USA  99, 9942-9947, US 2003-0166282, 2003-0143204, 2004-0038278, and 2003-0224432. 
         [0543]    Anti-sense agents can also be used to inhibit expression of a protein involved in GDP-fucose biosynthesis or a fucosyltransferase and include, for example, from about 8 to about 80 nucleobases (i.e. from about 8 to about 80 nucleotides), e.g., about 8 to about 50 nucleobases, or about 12 to about 30 nucleobases. Anti-sense compounds include ribozymes, external guide sequence (EGS) oligonucleotides (oligozymes), and other short catalytic RNAs or catalytic oligonucleotides that hybridize to the target nucleic acid and modulate its expression. Anti-sense compounds can include a stretch of at least eight consecutive nucleobases that are complementary to a sequence in the target gene. An oligonucleotide need not be 100% complementary to its target nucleic acid sequence to be specifically hybridizable. An oligonucleotide is specifically hybridizable when binding of the oligonucleotide to the target interferes with the normal function of the target molecule to cause a loss of utility, and there is a sufficient degree of complementarity to avoid non-specific binding of the oligonucleotide to non-target sequences under conditions in which specific binding is desired. 
         [0544]    Hybridization of antisense oligonucleotides with mRNA can interfere with one or more of the normal functions of mRNA. The functions of mRNA to be interfered with include all vital functions such as, for example, translocation of the RNA to the site of protein translation, translation of protein from the RNA, splicing of the RNA to yield one or more mRNA species, and catalytic activity that may be engaged in by the RNA. Binding of specific protein(s) to the RNA may also be interfered with by antisense oligonucleotide hybridization to the RNA. 
         [0545]    Exemplary antisense compounds include DNA or RNA sequences that specifically hybridize to the target nucleic acid. The complementary region can extend for between about 8 to about 80 nucleobases. The compounds can include one or more modified nucleobases. Modified nucleobases may include, e.g., 5-substituted pyrimidines such as 5-iodouracil, 5-iodocytosine, and C5-propynyl pyrimidines such as C5-propynylcytosine and C5-propynyluracil. Other suitable modified nucleobases include N4-(C1-C12)alkylaminocytosines and N4,N4-(C1-C12)dialkylaminocytosines. Modified nucleobases may also include 7-substituted-8-aza-7-deazapurines and 7-substituted-7-deazapurines such as, for example, 7-iodo-7-deazapurines, 7-cyano-7-deazapurines, 7-aminocarbonyl-7-deazapurines. Examples of these include 6-amino-7-iodo-7-deazapurines, 6-amino-7-cyano-7-deazapurines, 6-amino-7-aminocarbonyl-7-deazapurines, 2-amino-6-hydroxy-7-iodo-7-deazapurines, 2-amino-6-hydroxy-7-cyano-7-deazapurines, and 2-amino-6-hydroxy-7-aminocarbonyl-7-deazapurines. Furthermore, N6-(C1-C12)alkylaminopurines and N6,N6-(C1-C12)dialkylaminopurines, including N6-methylaminoadenine and N6,N6-dimethylaminoadenine, are also suitable modified nucleobases. Similarly, other 6-substituted purines including, for example, 6-thioguanine may constitute appropriate modified nucleobases. Other suitable nucleobases include 2-thiouracil, 8-bromoadenine, 8-bromoguanine, 2-fluoroadenine, and 2-fluoroguanine. Derivatives of any of the aforementioned modified nucleobases are also appropriate. Substituents of any of the preceding compounds may include C 1 -C 30  alkyl, C 2 -C 30  alkenyl, C 2 -C 30  alkynyl, aryl, aralkyl, heteroaryl, halo, amino, amido, nitro, thio, sulfonyl, carboxyl, alkoxy, alkylcarbonyl, alkoxycarbonyl, and the like. 
         [0546]    Descriptions of other types of nucleic acid agents are also available. See, e.g., U.S. Pat. No. 4,987,071; U.S. Pat. No. 5,116,742; U.S. Pat. No. 5,093,246; Woolf et al. (1992)  Proc Natl Acad Sci USA ; Antisense RNA and DNA, D. A. Melton, Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1988); 89:7305-9; Haselhoff and Gerlach (1988)  Nature  334:585-59; Helene, C. (1991)  Anticancer Drug Des.  6:569-84; Helene (1992)  Ann. N.Y. Acad. Sci.  660:27-36; and Maher, L. J. (1992)  Bioassays  14:807-15. 
       Genetically Engineered Cells 
       [0547]    In some embodiments, a cell can be selected that has been genetically engineered for permanent or regulated inactivation (complete or partial) of a gene encoding a gene involved in GDP-fucose biosynthesis or a fucosyltransferase, or a protein involved in regulating GDP-fucose levels. For example, genes described herein can be inactivated. Permanent or regulated inactivation of gene expression can be achieved by targeting to a gene locus with a transfected plasmid DNA construct or a synthetic oligonucleotide. The plasmid construct or oligonucleotide can be designed to several forms. These include the following: 1) insertion of selectable marker genes or other sequences within an exon of the gene being inactivated; 2) insertion of exogenous sequences in regulatory regions of non-coding sequence; 3) deletion or replacement of regulatory and/or coding sequences; and, 4) alteration of a protein coding sequence by site specific mutagenesis. 
         [0548]    In the case of insertion of a selectable marker gene into a coding sequence, it is possible to create an in-frame fusion of an endogenous exon of the gene with the exon engineered to contain, for example, a selectable marker gene. In this way following successful targeting, the endogenous gene expresses a fusion mRNA (nucleic acid sequence plus selectable marker sequence). Moreover, the fusion mRNA would be unable to produce a functional translation product. 
         [0549]    In the case of insertion of DNA sequences into regulatory regions, the transcription of a gene can be reduced or silenced by disrupting the endogenous promoter region or any other regions in the 5′ untranslated region (5′ UTR) that is needed for transcription. Such regions include, for example, translational control regions and splice donors of introns. Secondly, a new regulatory sequence can be inserted upstream of the gene that would alter expression, e.g., eliminate expression, reduce expression, or render the gene subject to the control of extracellular factors. It would thus be possible to down-regulate or extinguish gene expression as desired for glycoprotein production. Moreover, a sequence that includes a selectable marker and a promoter can be used to disrupt expression of the endogenous sequence. Finally, all or part of the endogenous gene could be deleted by appropriate design of targeting substrates. 
         [0550]    Cells Genetically Engineered to Express a Component Involved in Regulating GDP-fucose levels 
         [0551]    Cells can be genetically engineered to express a component involved in regulation of GDP-fucose levels, e.g., a cell can be genetically engineered to overexpress a GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, a GDP-fucose transporter, and/or a fucosyltransferase. When cells are to be genetically modified for the purposes of expressing or overexpressing a component, the cells may be modified by conventional genetic engineering methods or by gene activation. 
         [0552]    According to conventional methods, a DNA molecule that contains cDNA or genomic DNA sequence encoding desired protein may be contained within an expression construct and transfected into primary, secondary, or immortalized cells by standard methods including, but not limited to, liposome-, polybrene-, or DEAE dextran-mediated transfection, electroporation, calcium phosphate precipitation, microinjection, or velocity driven microprojectiles (see, e.g., U.S. Pat. No. 6,048,729). 
         [0553]    Alternatively, one can use a system that delivers the genetic information by viral vector. Viruses known to be useful for gene transfer include adenoviruses, adeno associated virus, herpes virus, mumps virus, pollovirus, retroviruses, Sindbis virus, and vaccinia virus such as canary pox virus. 
         [0554]    Alternatively, the cells may be modified using a gene activation approach, for example, as described in U.S. Pat. No. 5,641,670; U.S. Pat. No. 5,733,761; U.S. Pat. No. 5,968,502; U.S. Pat. No. 6,200,778; U.S. Pat. No. 6,214,622; U.S. Pat. No. 6,063,630; U.S. Pat. No. 6,187,305; U.S. Pat. No. 6,270,989; and U.S. Pat. No. 6,242,218. 
         [0555]    Accordingly, the term “genetically engineered,” as used herein in reference to cells, is meant to encompass cells that express a particular gene product following introduction of a DNA molecule encoding the gene product and/or including regulatory elements that control expression of a coding sequence for the gene product. The DNA molecule may be introduced by gene targeting or homologous recombination, i.e., introduction of the DNA molecule at a particular genomic site. 
         [0556]    Methods of transfecting cells, and reagents such as promoters, markers, signal sequences that can be used for recombinant expression are known. 
         [0557]    A component involved in regulating levels of GDP-fucose, e.g., GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, a fucosyltransferase or a GDP-fucose transporter, can be placed under a selected form of control, e.g., inducible control. For example, a sequence encoding GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, a fucosyltransferase or a GDP-fucose transporter, can be placed under the control of a promoter or other control element that is responsive to an inducer (or inhibitor) of expression. Such systems allow the cell to be maintained under a variety of conditions, e.g., a condition wherein the gene, e.g., a gene encoding GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, a fucosyltransferase or a GDP-fucose transporter, is expressed or not expressed. This allows culture of the cell under a first condition, which provides glycoproteins having a first glycosylation state (e.g., fucosylated), or under a second condition, which provides glycoproteins having a second glycosylation state (e.g., lacking fucosylation). 
         [0558]    Cells can also be engineered to express a hybrid nucleic acid; that is, a nucleic acid comprising at least two segments which have been isolated from at least two different sources. As one example of manipulation of a cell with a hybrid nucleic acid, a mammalian cell having a manipulation may express a hybrid nucleic acid comprising a regulatory sequence, such as a promoter and/or terminator sequence, of mammalian cell origin, which is functionally linked to a coding sequence, which may be of origin from a different species, e.g., from a different mammal or non-mammalian. In this manner, for example, a cell may be manipulated so that it can be induced to express the coding sequence in response to a stimulus that does not naturally induce expression of the linked coding sequence. An example of such a system is the TET On/Off regulatory system. In the Tet-Off system, gene expression is turned on when tetracycline (Tc) or doxycycline (Dox; a Tc derivative) is removed from the culture medium. In contrast, expression is turned on in the Tet-On system by the addition of Dox. The Tet-On system is responsive only to Dox, not to Tc. Both systems permit gene expression to be tightly regulated in response to varying concentrations of Tc or Dox. 
         [0559]    Generally, one of ordinary skill can select promoters for a desired level of gene expression and place a selected gene under the control of such a promoter. The term promoter as used herein refers to a polynucleotide sequence which allows and controls the transcription of the genes or sequences functionally connected therewith. The sequences of promoters are deposited in databases such as GeneBank, and may be obtained as separate elements or elements cloned within polynucleotide sequences from commercial or individual sources. Exemplary types of promoters that can be used to express a desired gene of interest in eukaryotic cells (e.g., animal cells) include, but not limited to, constitutive and inducible promoters. 
         [0560]    The activity of promoters may vary from one another in their strength, for example, across different cell types. Promoters that are particularly suitable for high expression in eukaryotic cells (e.g., animal cells) include, but not limited to, cytomegalovirus (CMV) immediate-early promoter, simian virus 40 (SV40) immediate-early promoter, human elongation factor 1α (EF-1α) promoter, chicken β-Actin promoter coupled with CMV early enhancer (CAG promoter), adenovirus major late promoter, and Rous sarcoma virus (RSV) promoter. Promoters that are suitable for intermediate or weak expression in eukaryotic cells (e.g., animal cells) include, but not limited to, human Ubiquitin C (UbC) promoter, murine phosphoglycerate kinase-1 (PGK) promoter, and herpes simplex virus (HSV) thymidine kinase (TK) promoter. Comparisons of the strength of various constitutive and inducible promoters in ectopic gene expression are described in, e.g., Qin, J. Y. et al.,  PLoS ONE  2010, 5(5):e10611; Cheng, X. et al.,  Int. J. Radiat. Biol.  1995, 67(3):261-267; Foecking, M. K. et al.,  Gene  1986, 45(1):101-105; Davis, M. G. et al.,  Biotechnol. Biochem.  1988, 10(1):6-12; Liu, Z. et al.,  Anal. Biochem.  1997, 246(1):150-152; Wenger, R. H. et al.,  Anal. Biochem.  1994, 221(2):416-418; Kronman, C. et al,  Gene  1992, 121(2):295-304; Thompson, T. A. et al.,  In Vitro Cell Dev. Biol.  1993, 29A (2):165-170; Thompson, E. M. et al.,  Gene  1990, 96(2):257-262). One of ordinary skill can evaluate a particular combination of promoter, gene, and cell line to obtain the desired level of expression. 
         [0561]    As mentioned above, with inducible promoters the activity of the promoter may be regulated (e.g., reduced or increased) in response to a signal (e.g., chemical signal (e.g., tetracycline, steroids, metal) or physical signal (e.g., temperature)). One example of an inducible promoter is the tetracycline (tet) promoter. As mentioned above, the tet promoter contains tetracycline a operator sequence (tetO) which can be induced by a tetracycline-regulated transactivator protein (tTA). Exemplary tetracycline-regulated promoters are described in e.g., U.S. Pat. Nos. 5,851,796, 5,464,758, 5,650,298, 5,589,362, 5,654,168, 5,789,156, 5,814,618, 5,888,981, 6,004,941, 6,136,954 and 6,271,348. Exemplary steroid-regulated promoters are described in e.g., U.S. Pat. Nos. 5,512,483 and 6,379,945. Exemplary metal-regulated promoters are described in e.g., U.S. Pat. Nos. 4,579,821 and 4,601,978. Examples of other inducible promoters include the jun, fos and heat shock promoter (see also Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1989; Gossen, M. et al.,  Curr. Opinions Biotech.  1994, 5, 516-520). 
         [0562]    The promoters described herein can be functionally combined with one or more regulatory sequences to regulate (e.g., increase, decrease, optimize, repress, induce) the transcription activity in an expression cassette. For example, the promoter can be functionally linked to one or more enhancer sequences (e.g., a CMV or SV40 enhancer) to increase transcriptional activity, or one or more binding sites for transcription factors (e.g., Sp1, AP1) to up- or down-regulate transcriptional activity. In an embodiment, the regulatory sequence can be positioned in front of or behind the promoter. 
       Transcription Factors 
       [0563]    The expression of a gene which conditions the level of GDP-fucose can also be down regulated by reducing, e.g., eliminating, the expression of a transcription factor which positively controls expression of the gene. E.g., Arnt, ATF6, SREBP-1c, Lmo2, HNF-1A, GCNF-2, CUTL1, STAT3, POU2F1a or EsF-1 can be targeted to down regulate GDP-fucose synthetase. HFH-1, Gfi-1, c-Myb, POU2F2C, AREB6, AORalpha2, POU3F1, LUN-1, or PPAR-gamma2 can be targeted to down regulate fucose kinase. Evi-1, STAT1beta, GATA-3, POU2F1A, POU3F2 (N-Oct-5b), AREB6, N-Myc, CUTL1, HSFlshort, or C/EBPbeta can be targeted to down regulate GNDS. 
       Chemical Inhibitors of GMD, FX, Fucokinase, GFPP or GDP-Fucose Synthetase 
       [0564]    Enzyme inhibitors are molecules that bind to enzymes and decrease their activities. The binding of an inhibitor may stop a substrate from entering the enzyme active site and/or hinder the enzyme from catalyzing its reaction. Inhibitor binding may be either reversible or irreversible. Irreversible inhibitors usually react with the enzyme and change it chemically. These inhibitors modify key amino acid residues needed for enzyme activity. In contrast, reversible inhibitors bind non-covalently and different types of inhibition are produced depending on whether these inhibitors bind the enzyme, the enzyme-substrate complex, or both. 
         [0565]    In some embodiments, the addition of particular chemical reagents or inhibitors may be used to lower the levels of the GDP-fucose. These reagents or inhibitors may inhibit GMD, FX, fucokinase, GFPP, GDP-fucose synthetase, or enzymes involved in the biosynthesis of GDP-mannose. Examples of these inhibitors include, but are not limited to, guanosine-5′-O-(2-thiodiphosphate)-fucose, guanosine-5′-O-(2-thiodiphosphate)-mannose, pyridoxal-5′-phosphate, GDP-4-dehydro-6-L-deoxygalactose, GDP-L-fucose, guanosine diphosphate (GDP), guanosine monophosphate (GMP), GDP-D-glucose, p-chloromercuriphenylsulfonate EDTA and fucose. 
       Glycoproteins 
       [0566]    Glycoproteins that can be made by methods described herein include those in Table 1 below. 
         [0000]    
       
         
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Protein Product 
                 Reference Listed Drug 
               
               
                   
               
             
             
               
                 interferon gamma-1b 
                 Actimmune ® 
               
               
                 alteplase; tissue plasminogen activator 
                 Activase ®/Cathflo ® 
               
               
                 Recombinant antihemophilic factor 
                 Advate 
               
               
                 human albumin 
                 Albutein ® 
               
               
                 Laronidase 
                 Aldurazyme ® 
               
               
                 interferon alfa-N3, human leukocyte derived 
                 Alferon N ® 
               
               
                 human antihemophilic factor 
                 Alphanate ® 
               
               
                 virus-filtered human coagulation factor IX 
                 AlphaNine ® SD 
               
               
                 Alefacept; recombinant, dimeric fusion protein LFA3-Ig 
                 Amevive ® 
               
               
                 Bivalirudin 
                 Angiomax ® 
               
               
                 darbepoetin alfa 
                 Aranesp ™ 
               
               
                 Bevacizumab 
                 Avastin ™ 
               
               
                 interferon beta-1a; recombinant 
                 Avonex ® 
               
               
                 coagulation factor IX 
                 BeneFix ™ 
               
               
                 Interferon beta-1b 
                 Betaseron ® 
               
               
                 Tositumomab 
                 BEXXAR ® 
               
               
                 antihemophilic factor 
                 Bioclate ™ 
               
               
                 human growth hormone 
                 BioTropin ™ 
               
               
                 botulinum toxin type A 
                 BOTOX ® 
               
               
                 Alemtuzumab 
                 Campath ® 
               
               
                 acritumomab; technetium-99 labeled 
                 CEA-Scan ® 
               
               
                 alglucerase; modified form of beta-glucocerebrosidase 
                 Ceredase ® 
               
               
                 imiglucerase; recombinant form of beta-glucocerebrosidase 
                 Cerezyme ® 
               
               
                 crotalidae polyvalent immune Fab, ovine 
                 CroFab ™ 
               
               
                 digoxin immune fab [ovine] 
                 DigiFab ™ 
               
               
                 Rasburicase 
                 Elitek ® 
               
               
                 Etanercept 
                 ENBREL ® 
               
               
                 epoietin alfa 
                 Epogen ® 
               
               
                 Cetuximab 
                 Erbitux ™ 
               
               
                 algasidase beta 
                 Fabrazyme ® 
               
               
                 Urofollitropin 
                 Fertinex ™ 
               
               
                 follitropin beta 
                 Follistim ™ 
               
               
                 Teriparatide 
                 FORTEO ® 
               
               
                 human somatropin 
                 GenoTropin ® 
               
               
                 Glucagon 
                 GlucaGen ® 
               
               
                 follitropin alfa 
                 Gonal-F ® 
               
               
                 antihemophilic factor 
                 Helixate ® 
               
               
                 Antihemophilic Factor; Factor XIII 
                 HEMOFIL 
               
               
                 adefovir dipivoxil 
                 Hepsera ™ 
               
               
                 Trastuzumab 
                 Herceptin ® 
               
               
                 Insulin 
                 Humalog ® 
               
               
                 antihemophilic factor/von Willebrand factor complex-human 
                 Humate-P ® 
               
               
                 Somatotropin 
                 Humatrope ® 
               
               
                 Adalimumab 
                 HUMIRA ™ 
               
               
                 human insulin 
                 Humulin ® 
               
               
                 recombinant human hyaluronidase 
                 Hylenex ™ 
               
               
                 interferon alfacon-1 
                 Infergen ® 
               
               
                 eptifibatide 
                 Integrilin ™ 
               
               
                 alpha-interferon 
                 Intron A ® 
               
               
                 Palifermin 
                 Kepivance 
               
               
                 Anakinra 
                 Kineret ™ 
               
               
                 antihemophilic factor 
                 Kogenate ®FS 
               
               
                 insulin glargine 
                 Lantus ® 
               
               
                 granulocyte macrophage colony-stimulating factor 
                 Leukine ®/Leukine ® Liquid 
               
               
                 lutropin alfa for injection 
                 Luveris 
               
               
                 OspA lipoprotein 
                 LYMErix ™ 
               
               
                 Ranibizumab 
                 LUCENTIS ® 
               
               
                 gemtuzumab ozogamicin 
                 Mylotarg ™ 
               
               
                 Galsulfase 
                 Naglazyme ™ 
               
               
                 Nesiritide 
                 Natrecor ® 
               
               
                 Pegfilgrastim 
                 Neulasta ™ 
               
               
                 Oprelvekin 
                 Neumega ® 
               
               
                 Filgrastim 
                 Neupogen ® 
               
               
                 Fanolesomab 
                 NeutroSpec ™ (formerly LeuTech ®) 
               
               
                 somatropin [rDNA] 
                 Norditropin ®/Norditropin Nordiflex ® 
               
               
                 Mitoxantrone 
                 Novantrone ® 
               
               
                 insulin; zinc suspension; 
                 Novolin L ® 
               
               
                 insulin; isophane suspension 
                 Novolin N ® 
               
               
                 insulin, regular; 
                 Novolin R ® 
               
               
                 Insulin 
                 Novolin ® 
               
               
                 coagulation factor VIIa 
                 NovoSeven ® 
               
               
                 Somatropin 
                 Nutropin ® 
               
               
                 immunoglobulin intravenous 
                 Octagam ® 
               
               
                 PEG-L-asparaginase 
                 Oncaspar ® 
               
               
                 abatacept, fully human soluable fusion protein 
                 Orencia ™ 
               
               
                 muromomab-CD3 
                 Orthoclone OKT3 ® 
               
               
                 high-molecular weight hyaluronan 
                 Orthovisc ® 
               
               
                 human chorionic gonadotropin 
                 Ovidrel ® 
               
               
                 live attenuated Bacillus Calmette-Guerin 
                 Pacis ® 
               
               
                 peginterferon alfa-2a 
                 Pegasys ® 
               
               
                 pegylated version of interferon alfa-2b 
                 PEG-Intron ™ 
               
               
                 Abarelix (injectable suspension); gonadotropin-releasing 
                 Plenaxis ™ 
               
               
                 hormone antagonist 
               
               
                 epoietin alfa 
                 Procrit ® 
               
               
                 Aldesleukin 
                 Proleukin, IL-2 ® 
               
               
                 Somatrem 
                 Protropin ® 
               
               
                 dornase alfa 
                 Pulmozyme ® 
               
               
                 Efalizumab; selective, reversible T-cell blocker 
                 RAPTIVA ™ 
               
               
                 combination of ribavirin and alpha interferon 
                 Rebetron ™ 
               
               
                 Interferon beta 1a 
                 Rebif ® 
               
               
                 antihemophilic factor 
                 Recombinate ® rAHF/ 
               
               
                 antihemophilic factor 
                 ReFacto ® 
               
               
                 Lepirudin 
                 Refludan ® 
               
               
                 Infliximab 
                 REMICADE ® 
               
               
                 Abciximab 
                 ReoPro ™ 
               
               
                 Reteplase 
                 Retavase ™ 
               
               
                 Rituxima 
                 Rituxan ™ 
               
               
                 interferon alfa-2 a   
                 Roferon-A ® 
               
               
                 Somatropin 
                 Saizen ® 
               
               
                 synthetic porcine secretin 
                 SecreFlo ™ 
               
               
                 Basiliximab 
                 Simulect ® 
               
               
                 Eculizumab 
                 SOLIRIS (R) 
               
               
                 Pegvisomant 
                 SOMAVERT ® 
               
               
                 Palivizumab; recombinantly produced, humanized mAb 
                 Synagis ™ 
               
               
                 thyrotropin alfa 
                 Thyrogen ® 
               
               
                 Tenecteplase 
                 TNKase ™ 
               
               
                 Natalizumab 
                 TYSABRI ® 
               
               
                 human immune globulin intravenous 5% and 10% solutions 
                 Venoglobulin-S ® 
               
               
                 interferon alfa-n1, lymphoblastoid 
                 Wellferon ® 
               
               
                 drotrecogin alfa 
                 Xigris ™ 
               
               
                 Omalizumab; recombinant DNA-derived humanized 
                 Xolair ® 
               
               
                 monoclonal antibody targeting immunoglobulin-E 
               
               
                 Daclizumab 
                 Zenapax ® 
               
               
                 ibritumomab tiuxetan 
                 Zevalin ™ 
               
               
                 Somatotropin 
                 Zorbtive ™ (Serostim ®) 
               
               
                   
               
             
          
         
       
     
       Analytical Methods 
       [0567]    In general, a glycan preparation can be subjected to analysis to determine whether the glycan includes a particular type of structure (e.g., a glycan structure described herein). In some embodiments, the analysis comprises comparing the structure and/or function of glycans in one glycoprotein preparation to structure and/or function of glycans in at least one other glycoprotein preparation. In some embodiments, the analysis comprises comparing the structure and/or function of glycans in one or more of the samples to structure and/or function of glycans in a reference sample. 
         [0568]    Structure and composition of glycans can be analyzed by any available method. In some embodiments, glycan structure and composition are analyzed by chromatographic methods, mass spectrometry (MS) methods, chromatographic methods followed by MS, electrophoretic methods, electrophoretic methods followed by MS, nuclear magnetic resonance (NMR) methods, and combinations thereof. 
         [0569]    In some embodiments, glycan structure and composition can be analyzed by chromatographic methods, including but not limited to, liquid chromatography (LC), high performance liquid chromatography (HPLC), ultra performance liquid chromatography (HPLC), thin layer chromatography (TLC), amide column chromatography, and combinations thereof. 
         [0570]    In some embodiments, glycan structure and composition can be analyzed by mass spectrometry (MS) and related methods, including but not limited to, tandem MS, LC-MS, LC-MS/MS, matrix assisted laser desorption ionisation mass spectrometry (MALDI-MS), Fourier transform mass spectrometry (FTMS), ion mobility separation with mass spectrometry (IMS-MS), electron transfer dissociation (ETD-MS), and combinations thereof. 
         [0571]    In some embodiments, glycan structure and composition can be analyzed by electrophoretic methods, including but not limited to, capillary electrophoresis (CE), CE-MS, gel electrophoresis, agarose gel electrophoresis, acrylamide gel electrophoresis, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting using antibodies that recognize specific glycan structures, and combinations thereof. 
         [0572]    In some embodiments, glycan structure and composition can be analyzed by nuclear magnetic resonance (NMR) and related methods, including but not limited to, one-dimensional NMR (1D-NMR), two-dimensional NMR (2D-NMR), correlation spectroscopy magnetic-angle spinning NMR (COSY-NMR), total correlated spectroscopy NMR (TOCSY-NMR), heteronuclear single-quantum coherence NMR (HSQC-NMR), heteronuclear multiple quantum coherence (HMQC-NMR), rotational nuclear overhauser effect spectroscopy NMR (ROESY-NMR), nuclear overhauser effect spectroscopy (NOESY-NMR), and combinations thereof. 
         [0573]    In some embodiments, techniques described herein may be combined with one or more other technologies for the detection, analysis, and or isolation of glycans or glycoproteins. For example, in certain embodiments, glycans are analyzed in accordance with the present disclosure using one or more available methods (to give but a few examples, see Anumula,  Anal. Biochem.  350(1):1, 2006; Klein et al.,  Anal. Biochem.,  179:162, 1989; and/or Townsend, R.R. Carbohydrate Analysis” High Performance Liquid Chromatography and Capillary Electrophoresis, Ed. Z. El Rassi, pp 181-209, 1995, each of which is incorporated herein by reference in its entirety). For example, in some embodiments, glycans are characterized using one or more of chromatographic methods, electrophoretic methods, nuclear magnetic resonance methods, and combinations thereof. Exemplary such methods include, for example, NMR, mass spectrometry, liquid chromatography, 2-dimensional chromatography, SDS-PAGE, antibody staining, lectin staining, monosaccharide quantitation, capillary electrophoresis, fluorophore-assisted carbohydrate electrophoresis (FACE), micellar electrokinetic chromatography (MEKC), exoglycosidase or endoglycosidase treatments, and combinations thereof. Those of ordinary skill in the art will be aware of other techniques that can be used to characterize glycans together with the methods described herein. 
         [0574]    In some embodiments, methods described herein allow for detection of a glycan structure (such as a glycan structure described herein) that is present at low levels within a population of glycans. For example, the present methods allow for detection of glycan species that are present at levels less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.5%, less than 1%, less than 0.75%, less than 0.5%, less than 0.25%, less than 0.1%, less than 0.075%, less than 0.05%, less than 0.025%, or less than 0.01% within a population of glycans. 
         [0575]    In some embodiments, methods described herein allow for detection of particular structures (e.g., a glycan structure described herein) that are present at low levels within a population of glycans. For example, the present methods allow for detection of particular structures that are present at levels less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.5%, less than 1%, less than 0.75%, less than 0.5%, less than 0.25%, less than 0.1%, less than 0.075%, less than 0.05%, less than 0.025%, or less than 0.01% within a population of glycans. 
         [0576]    In some embodiments, methods described herein allow for detection of relative levels of individual glycan species within a population of glycans. For example, the area under each peak of a liquid chromatograph can be measured and expressed as a percentage of the total. Such an analysis provides a relative percent amount of each glycan species within a population of glycans. In another example, relative levels of individual glycan species are determined from areas of peaks in a 1D-NMR experiment, or from volumes of cross peaks from a  1 H- 15 N HSQC spectrum (e.g., with correction based on responses from standards), or by relative quantitation by comparing the same peak across samples. 
         [0577]    In some embodiments, a biological activity of a glycoprotein preparation (e.g., a glycoprotein preparation) is assessed. Biological activity of glycoprotein preparations can be analyzed by any available method. In some embodiments, a binding activity of a glycoprotein is assessed (e.g., binding to a receptor). In some embodiments, a therapeutic activity of a glycoprotein is assessed (e.g., an activity of a glycoprotein in decreasing severity or symptom of a disease or condition, or in delaying appearance of a symptom of a disease or condition). In some embodiments, a pharmacologic activity of a glycoprotein is assessed (e.g., bioavailability, pharmacokinetics, pharmacodynamics). For methods of analyzing bioavailability, pharmacokinetics, and pharmacodynamics of glycoprotein therapeutics, see, e.g., Weiner et al.,  J. Pharm. Biomed. Anal.  15(5):571-9, 1997; Srinivas et al.,  J. Pharm. Sci.  85(1):1-4, 1996; and Srinivas et al.,  Pharm. Res.  14(7):911-6, 1997. 
         [0578]    As would be understood to one of skill in the art, the particular biological activity or therapeutic activity that can be tested will vary depending on the particular glycoprotein or glycan structure. 
         [0579]    The potential adverse activity or toxicity (e.g., propensity to cause hypertension, allergic reactions, thrombotic events, seizures, or other adverse events) of glycoprotein preparations can be analyzed by any available method. In some embodiments, immunogenicity of a glycoprotein preparation is assessed, e.g., by determining whether the preparation elicits an antibody response in a subject. 
       Cells &amp; Cell Lines 
       [0580]    Methods described herein use cells to produce products having reduced fucosylation. Examples of cells useful in these and other methods described herein follow. 
         [0581]    The cell useful in the methods described herein can be eukaryotic or prokaryotic, as long as the cell provides or has added to it the enzymes to activate and attach saccharides present in the cell or saccharides present in the cell culture medium or fed to the cells. Examples of eukaryotic cells include yeast, insect, fungi, plant and animal cells, especially mammalian cells. Suitable mammalian cells include any normal mortal or normal or abnormal immortal animal or human cell, including: monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293) (Graham et al., J. Gen. Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese Hamster Ovary (CHO), e.g., DG44, DUKX-V11, GS-CHO (ATCC CCL 61, CRL 9096, CRL 1793 and CRL 9618); mouse sertoli cells (TM4, Mather, Biol. Reprod. 23:243 251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL 1587); human cervical carcinoma cells (HeLa, ATCC CCL 2); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse melanoma cells (NSO); mouse mammary tumor (MMT 060562, ATCC CCL51), TR1 cells (Mather, et al., Annals N.Y. Acad. Sci. 383:44 46 (1982)); canine kidney cells (MDCK) (ATCC CCL 34 and CRL 6253), HEK 293 (ATCC CRL 1573), WI-38 cells (ATCC CCL 75) (ATCC: American Type Culture Collection, Rockville, Md.), MCF-7 cells, MDA-MB-438 cells, U87 cells, A127 cells, HL60 cells, A549 cells, SP10 cells, DOX cells, SHSY5Y cells, Jurkat cells, BCP-1 cells, GH3 cells, 9L cells, MC3T3 cells, C3H-10T1/2 cells, NIH-3T3 cells, C6/36 cells, human lymphoblast cell lines (e.g. GEX) and PER.C6® cells. The use of mammalian tissue cell culture to express polypeptides is discussed generally in Winnacker, FROM GENES TO CLONES (VCH Publishers, N.Y., N.Y., 1987). 
         [0582]    Exemplary plant cells include, for example,  Arabidopsis thaliana , rape seed, corn, wheat, rice, tobacco etc.) (Staub, et al. 2000 Nature Biotechnology 1(3): 333-338 and McGarvey, P. B., et al. 1995 Bio-Technology 13(13): 1484-1487; Bardor, M., et al. 1999 Trends in Plant Science 4(9): 376-380). Exemplary insect cells (for example,  Spodoptera frugiperda  Sf9, Sf21,  Trichoplusia ni , etc. Exemplary bacteria cells include  Escherichia coli . Various yeasts and fungi such as  Pichia pastoris, Pichia methanolica, Hansenula polymorpha , and  Saccharomyces cerevisiae  can also be selected. 
         [0583]    Culture Media and Processing 
         [0584]    The methods described herein can include determining and/or selecting media components or culture conditions which result in the production of a desired glycan property or properties. Culture parameters that can be determined include media components, pH, feeding conditions, osmolarity, carbon dioxide levels, agitation rate, temperature, cell density, seeding density, timing and sparge rate. 
         [0585]    Changes in production parameters such the speed of agitation of a cell culture, the temperature at which cells are cultures, the components in the culture medium, the times at which cultures are started and stopped, variation in the timing of nutrient supply can result in variation of a glycan properties of the produced glycoprotein product. Thus, methods described herein can include one or more of: increasing or decreasing the speed at which cells are agitated, increasing or decreasing the temperature at which cells are cultures, adding or removing media components, and altering the times at which cultures are started and/or stopped. 
         [0586]    Sequentially selecting a production parameters or a combination thereof, as used herein, means a first parameter (or combination) is selected, and then a second parameter (or combination) is selected, e.g., based on a constraint imposed by the choice of the first production parameter. 
         [0587]    Media 
         [0588]    The methods described herein can include determining and/or selecting a media component and/or the concentration of a media component that has a positive correlation to a desired glycan property or properties. A media component can be added in or administered over the course of glycoprotein production or when there is a change in media, depending on culture conditions. Media components include components added directly to culture as well as components that are a byproduct of cell culture. 
         [0589]    Media components include, e.g., buffer, amino acid content, vitamin content, salt content, mineral content, serum content, carbon source content, lipid content, nucleic acid content, hormone content, trace element content, ammonia content, co-factor content, indicator content, small molecule content, hydrolysate content and enzyme modulator content. Specific examples of media conditions that will lead to altered levels of GDP-fucose include but are not limited to altering the levels of cobalt, butyrate, fucose, guanosine, and manganese. 
         [0590]    Table 2 provides examples of various media components that can be selected. 
         [0000]    
       
         
               
               
             
           
               
                 TABLE 2 
               
               
                   
               
             
             
               
                 amino acids 
                 sugar precursors 
               
               
                 Vitamins 
                 Indicators 
               
               
                 Carbon source (natural and unnatural) 
                 Nucleosides or nucleotides 
               
               
                 Salts 
                 butyrate or organics 
               
               
                 Sugars 
                 DMSO 
               
               
                 Sera 
                 Animal derived products 
               
               
                 Plant derived hydrolysates 
                 Gene inducers 
               
               
                 sodium pyruvate 
                 Non natural sugars 
               
               
                 Surfactants 
                 Regulators of intracellular pH 
               
               
                 Ammonia 
                 Betaine or osmoprotectant 
               
               
                 Lipids 
                 Trace elements 
               
               
                 Hormones or growth factors 
                 minerals 
               
               
                 Buffers 
                 Non natural amino acids 
               
               
                 Non natural amino acids 
                 Non natural vitamins 
               
               
                   
               
             
          
         
       
     
         [0591]    Exemplary buffers include Tris, Tricine, HEPES, MOPS, PIPES, TAPS, bicine, BES, TES, cacodylate, MES, acetate, MKP, ADA, ACES, glycinamide and acetamidoglycine. 
         [0592]    The media can be serum free or can include animal derived products such as, e.g., fetal bovine serum (FBS), fetal calf serum (FCS), horse serum (HS), human serum, animal derived serum substitutes (e.g., Ultroser G, SF and HY; non-fat dry milk; Bovine EX-CYTE), fetuin, bovine serum albumin (BSA), serum albumin, and transferrin. When serum free media is selected lipids such as, e.g., palmitic acid and/or steric acid, can be included. 
         [0593]    Lipids components include oils, saturated fatty acids, unsaturated fatty acids, glycerides, steroids, phospholipids, sphingolipids and lipoproteins. 
         [0594]    Exemplary amino acid that can be included or eliminated from the media include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, proline, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine. 
         [0595]    Examples of vitamins that can be present in the media or eliminated from the media include vitamin A (retinoid), vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6 (pyroxidone), vitamin B7 (biotin), vitamin B9 (folic acid), vitamin B12 (cyanocobalamin), vitamin C (ascorbic acid), vitamin D, vitamin E, and vitamin K. 
         [0596]    Minerals that can be present in the media or eliminated from the media include bismuth, boron, calcium, chlorine, chromium, cobalt, copper, fluorine, iodine, iron, magnesium, manganese, molybdenum, nickel, phosphorus, potassium, rubidium, selenium, silicon, sodium, strontium, sulfur, tellurium, titanium, tungsten, vanadium, and zinc. Exemplary salts and minerals include CaCl 2  (anhydrous), CuSO 4  5H 2 O, Fe(NO 3 ).9H 2 O, KCl, KNO 3 , KH 2 PO 4 , MgSO 4  (anhydrous), NaCl, NaH 2 PO 4 H 2 O, NaHCO 3 , Na 2 SeO 3  (anhydrous), ZnSO 4 .7H 2 O; linoleic acid, lipoic acid, D-glucose, hypoxanthine 2Na, phenol red, putrescine 2HCl, sodium pyruvate, thymidine, pyruvic acid, sodium succinate, succinic acid, succinic acid.Na.hexahydrate, glutathione (reduced), para-aminobenzoic acid (PABA), methyl linoleate, bacto peptone G, adenosine, cytidine, guanosine, 2′-deoxyadenosine HCl, 2′-deoxycytidine HCl, 2′-deoxyguanosine and uridine. When the desired glycan characteristic is decreased fucosylation, the production parameters can include culturing a cell, e.g., CHO cell, e.g., dhfr deficient CHO cell, in the presence of manganese, e.g., manganese present at a concentration of about 0.1 μM to 50 μM. Decreased fucosylation can also be obtained, e.g., by culturing a cell (e.g., a CHO cell, e.g., a dhfr deficient CHO cell) at an osmolality of about 350 to 500 mOsm. Osmolality can be adjusted by adding salt to the media or having salt be produced as a byproduct as evaporation occurs during production. 
         [0597]    Hormones include, for example, somatostatin, growth hormone-releasing factor (GRF), insulin, prolactin, human growth hormone (hGH), somatotropin, estradiol, and progesterone. Growth factors include, for example, bone morphogenic protein (BMP), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), nerve growth factor (NGF), bone derived growth factor (BDGF), transforming growth factor-beta1 (TGF-beta1), [Growth factors from U.S. Pat. No. 6,838,284 B2], hemin and NAD. 
         [0598]    Examples of surfactants that can be present or eliminated from the media include Tween-80 and pluronic F-68. 
         [0599]    Small molecules can include, e.g., butyrate, ammonia, non natural sugars, non natural amino acids, chloroquine, and betaine. 
         [0600]    In some embodiments, ammonia content can be selected as a production parameter to produce a desired glycan characteristic or characteristics. For example, ammonia can be present in the media in a range from 0.001 to 50 mM. Ammonia can be directly added to the culture and/or can be produced as a by product of glutamine or glucosamine. When the desired glycan characteristic is one or more of an increased number of high mannose structures, decreased fucosylation and decreased galactosylation, the production parameters selected can include culturing a cell (e.g., a CHO cell, e.g., a dhfr deficient CHO cell) in the presence of ammonia, e.g., ammonia present at a concentration of about 0.01 to 50 mM. For example, if the desired glycan characteristic includes decreased galactosylation, production parameters selected can include culturing a cell (e.g., a CHO cell, e.g., a dhfr deficient CHO cell) in serum containing media and in the presence of ammonia, e.g., ammonia present at a concentration of about 0.01 to 50 mM. 
         [0601]    Another production parameter is butyrate content. The presence of butyrate in culture media can result in increased galactose levels in the resulting glycoprotein preparation. Butyrate provides increased sialic acid content in the resulting glycoprotein preparation. Therefore, when increased galactosylation and/or sialylation is desired, the cell used to produce the glycoprotein (e.g., a CHO cell, e.g., a dhfr deficient CHO cell) can be cultured in the presence of butyrate. In some embodiments, butyrate can be present at a concentration of about 0.001 to 10 mM, e.g., about 2 mM to 10 mM. For example, if the desired glycan characteristic includes increased sialylation, production parameters selected can include culturing a cell (e.g., a CHO cell, e.g., a dhfr deficient CHO cell) in serum containing media and in the presence of butyrate, e.g., butyrate present at a concentration of about 2.0 to 10 mM. Such methods can further include selecting one or more of adherent culture conditions and culture in a T flask. 
         [0602]    Physiochemical Parameters 
         [0603]    Methods described herein can include selecting culture conditions that are correlated with a desired glycan property or properties. Such conditions can include temperature, pH, osmolality, shear force or agitation rate, oxidation, spurge rate, growth vessel, tangential flow, DO, CO 2 , nitrogen, fed batch, redox, cell density and feed strategy. Examples of physiochemical parameters that can be selected are provided in Table 3. 
         [0000]    
       
         
               
               
               
             
           
               
                   
                 TABLE 3 
               
               
                   
                   
               
             
             
               
                   
                 Temperature 
                 DO 
               
               
                   
                 pH 
                 CO 2   
               
               
                   
                 osmolality 
                 Nitrogen 
               
               
                   
                 shear force, or agitation rate 
                 Fed batch 
               
               
                   
                 oxidation 
                 Redox 
               
               
                   
                 Spurge rate 
                 Cell density 
               
               
                   
                 Growth vessel 
                 Perfusion culture 
               
               
                   
                 Tangential flow 
                 Feed strategy 
               
               
                   
                 Batch 
               
               
                   
                   
               
             
          
         
       
     
         [0604]    For example, the production parameter can be culturing a cell under acidic, neutral or basic pH conditions. Temperatures can be selected from 10 to 42° C. For example, a temperature of about 28 to 36° C. does not significantly alter galactosylation, fucosylation, high mannose production, hybrid production or sialylation of glycoproteins produced by a cell (e.g., a CHO cell, e.g., a dhfr deficient CHO cell) cultured at these temperatures. In addition, any method that slows down the growth rate of a cell may also have this effect. Thus, temperatures in this range or methods that slow down growth rate can be selected when it is desirable not to have this parameter of production altering glycosynthesis. 
         [0605]    In other embodiments, carbon dioxide levels can be selected which results in a desired glycan characteristic or characteristics. CO 2  levels can be, e.g., about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 13%, 15%, 17%, 20%, 23% and 25% (and ranges in between). In one embodiment, when decreased fucosylation is desired, the cell can be cultured at CO 2  levels of about 11 to 25%, e.g., about 15%. CO 2  levels can be adjusted manually or can be a cell byproduct. 
         [0606]    A wide array of flasks, bottles, reactors, and controllers allow the production and scale up of cell culture systems. The system can be chosen based, at least in part, upon its correlation with a desired glycan property or properties. 
         [0607]    Cells can be grown, for example, as batch, fed-batch, perfusion, or continuous cultures. 
         [0608]    Production parameters that can be selected include, e.g., addition or removal of media including when (early, middle or late during culture time) and how often media is harvested; increasing or decreasing speed at which cell cultures are agitated; increasing or decreasing temperature at which cells are cultured; adding or removing media such that culture density is adjusted; selecting a time at which cell cultures are started or stopped; and selecting a time at which cell culture parameters are changed. Such parameters can be selected for any of the batch, fed-batch, perfusion and continuous culture conditions. 
       EXAMPLES 
     Example 1 
     Relationship Between Levels of GDP-Fucose and % Fucosylated Glycans 
       [0609]    The levels of GDP-fucose levels and the degree of protein fucosylation on glycoproteins were analyzed for three different CHO cell lines expressing a representative secreted protein product (CTLA4Ig): CHO cells that are deficient in the enzyme GDP-mannose 4,6, dehydratase (Lec 13.6 A); CHO cells that have lowered levels of GDP-fucose (Lec 2); and wild-type CHO cells. Culture media did not contain free fucose except as indicated for Lec 13.6 A cells cultured in the presence of exogenous fucose supplemented at 0.01 and 1 mM in the culture media. Cells were harvested, and snap frozen, while culture supernatant was harvested and CTLA4Ig harvested by protein A purification for subsequent analysis. Cells were then subjected to nucleotide sugar extraction using standard methods. In short with chloroform:methanol:water (2:4:1), the pellets discarded and the resulting extraction dried down. The dried material was subsequently resuspended in 500 ul of 10% butanol in water and then extracted with 1 ml of 90% butanol in water. The butanol phase was discarded and the aqueous subjected to a second butanol extraction. The final aqueous phase was dried down and the sugar nucleotides further isolated by PGC chromatography eluting off with 25% acetonitrile (v/v) containing 50 mM triethylammonium acetate. For quantification, sugar-nucletides were resolved with RP chromatography. 
         [0610]    Protein products were isolated from culture supernatant by protein A affinity, and subjected to PNGase F treatment to remove glycans. The resulting glycans were isolated by PGC chromatography and subsequently analyzed by MALDI mass spectrometry. The % fucosylation was determined by determining the ratio of the glycans with or without core fucosylation. Results are presented in Table 4. GDP-fucose levels are indicated in peak area as detected by UV. 
         [0000]    
       
         
               
               
               
             
               
               
               
             
           
               
                 TABLE 4 
               
               
                   
               
               
                   
                 % of Parental 
                 % Fucosylated 
               
               
                 Cell Line 
                 GDP-fucose 
                 Glycans 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Wild-type CHO 
                 100 
                 &gt;90 
               
               
                 Lec 2 
                 80 
                 &gt;90 
               
               
                 Lec 13.6A 
                 61.6 
                 ≈20 
               
               
                 Lec 13.6A + 1 mM fucose 
                 270 
                 100 
               
               
                 Lec 13.6A + 0.01 mM fucose 
                 62.5 
                 45

Technology Classification (CPC): 2