Patent Publication Number: US-2011076270-A1

Title: Therapeutic Binding Molecules

Description:
FIELD OF THE INVENTION 
     The present invention relates to organic compounds, such as to binding molecules against CD45 antigen isoforms, such as for example monoclonal antibodies (mAbs). 
     BACKGROUND OF THE INVENTION 
     One approach in the treatment of a variety of diseases is to achieve the elimination or the inactivation of pathogenic leukocytes and the potential for induction of tolerance to inactivate pathological immune responses. 
     Organ, cell and tissue transplant rejection and the various autoimmune diseases are thought to be primarily the result of T-cell mediated immune response triggered by helper T-cells which are capable of recognizing specific antigens which are captured, processed and presented to the helper T cells by antigen presenting cell (APC) such as macrophages and dendritic cells, in the form of an antigen-MHC complex, i.e. the helper T-cell when recognizing specific antigens is stimulated to produce cytokines such as IL-2 and to express or upregulate some cytokine receptors and other activation molecules and to proliferate. Some of these activated helper T-cells may act directly or indirectly, i.e. assisting effector cytotoxic T-cells or B cells, to destroy cells or tissues expressing the selected antigen. After the termination of the immune response some of the mature clonally selected cells remain as memory helper and memory cytotoxic T-cells, which circulate in the body and rapidly recognize the antigen if appearing again. If the antigen triggering this response is an innocuous environmental antigen the result is allergy, if the antigen is not a foreign antigen, but a self antigen, it can result is autoimmune disease; if the antigen is an antigen from a transplanted organ, the result can be graft rejection. 
     The immune system has developed to recognize self from non-self. This property enables an organism to survive in an environment exposed to the daily challenges of pathogens. This specificity for non-self and tolerance towards self arises during the development of the T cell repertoire in the thymus through processes of positive and negative selection, which also comprise the recognition and elimination of autoreactive T cells. This type of tolerance is referred to as central tolerance. However, some of these autoreactive cells escape this selective mechanism and pose a potential hazard for the development of autoimmune diseases. To control the autoreactive T cells that have escaped to the periphery, the immune system has peripheral regulatory mechanisms that provide protection against autoimmunity. These mechanisms are a basis for peripheral tolerance. 
     Cell surface antigens recognized by specific mAbs are generally designated by a CD (Cluster of Differentiation) number assigned by successive International Leukocyte Typing workshops and the term CD45 applied herein refers to the cell surface leukocyte common antigen CD45; and an mAb to that antigen is designated herein as “anti-CD45”. 
     Antibodies against the leukocyte common antigen (LCA) or CD45 are a major component of anti-lymphocyte globulin (ALG). CD45 belongs to the family of transmembrane tyrosine phosphatases and is both a positive and negative regulator of cell activation, depending upon receptor interaction. The phosphatase activity of CD45 appears to be required for activation of Src-family kinases associated with antigen receptor of B and T lymphocytes (Trowbridge I S et al, Annu Rev Immunol. 1994; 12:85-116). Thus, in T cell activation, CD45 is essential for signal 1 and CD45-deficient cells have profound defects in TCR-mediated activation events. 
     The CD45 antigen exists in different isoforms comprising a family of transmembrane glycoproteins. Distinct isoforms of CD45 differ in their extracellular domain structure which arise from alternative splicing of 3 variable exons coding for part of the CD45 extracellular region (Streuli M F. et al, J. Exp. Med. 1987; 166:1548-1566). The various isoforms of CD45 have different extra-cellular domains, but have the same transmembrane and cytoplasmic segments having two homologous, highly conserved phosphatase domains of approximately 300 residues. Different isoform combinations are differentially expressed on subpopulations of T and B lymphocytes (Thomas M L. et al, Immunol. Today 1988; 9:320-325). Some monoclonal antibodies recognize an epitope common to all the different isoforms, while other mAbs have a restricted (CD45R) specificity, dependent on which of the alternatively spliced exons (A, B or C) they recognize. For example, monoclonal antibodies recognizing the product of exon A are consequently designated CD45RA, those recognizing the various isoforms containing exon B have been designated CD45RB (Beverley P C L et al, Immunol. Supp. 1988; I:3-5). Antibodies such as UCHL1 selectively bind to the 180 kDa isoform CD45RO (without any of the variable exons A, B or C) which appears to be restricted to a subset of activated T cells, memory cells and cortical thymocytes and is not detected on B cells (Terry L A et al, Immunol. 1988; 64:331-336). 
    
    
     
       DESCRIPTION OF THE FIGURES 
         FIG. 1  shows that the inhibition of primary MLR by the “candidate mAb” is dose-dependent in the range of 0.001 and 10 μg/ml. “Concentration” is concentration of the “candidate mAb”. 
         FIG. 2  shows the plasmid map of the expression vector HCMV-G1 HuAb-VHQ comprising the heavy chain having the nucleotide sequence SEQ ID NO:12 (3921-4274) in the complete expression vector nucleotide sequence SEQ ID NO:15. 
         FIG. 3  shows the plasmid map of the expression vector HCMV-G1 HuAb-VHE comprising the heavy chain having the nucleotide sequence SEQ ID NO:11 (3921-4274) in the complete expression vector nucleotide sequence SEQ ID NO:16. 
         FIG. 4  shows the plasmid map of the expression vector HCMV-K HuAb-humV1 comprising the light chain having the nucleotide sequence SEQ ID NO:14 (3964-4284) in the complete expression vector nucleotide sequence SEQ ID NO:17. 
         FIG. 5  shows the plasmid map of the expression vector HCMV-K HuAb-humV2 comprising the light chain having the nucleotide sequence SEQ ID NO:13 (3926-4246) in the complete expression vector nucleotide sequence SEQ ID NO:18. 
     
    
    
     DESCRIPTION OF THE INVENTION 
     We have now found a binding molecule which comprises a polypeptide sequence which binds to CD45RO and CD45RB, hereinafter also designated as a “CD45RO/RB binding molecule”. These binding molecule according to the invention may induce immunosuppression, inhibit primary T cell responses and induce T cell tolerance. Furthermore, the binding molecules of the invention inhibit primary mixed lymphocyte responses (MLR). Cells derived from cultures treated with CD45RO/RB binding molecules preferredly also have impaired proliferative responses in secondary MLR even in the absence of CD45RO/RB binding molecules in the secondary MLR. Such impaired proliferative responses in secondary MLR are an indication of the ability of binding molecules of the invention to induce tolerance. 
     Furthermore, it is found that in vivo administration of CD45RO/RB binding molecule to severe combined immunodeficiency (SCID) mice undergoing xeno-GVHD following injection with human PBMC may prolong mice survival, compared to control treated mice, even though circulating human T cells may still be detected in CD45RO/RB binding molecule treated mice. CD45RB/RO binding molecule may also suppress the inflammatory process that mediates human allograft skin rejection. 
     By “CD45RO/RB binding molecule” is meant any molecule capable of binding specifically to the CD45RB and CD45RO isoforms of the CD45 antigen, either alone or associated with other molecules. The binding reaction may be shown by standard methods (qualitative assay) including for example any kind of binding assay such as direct or indirect immunofluorescence together with fluorescence microscopy or cytofluorimetric (FACS) analysis, enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay in which binding of the molecule to cells expressing a particular CD45 isoform can be visualized. In addition, the binding of this molecule may result in the alteration of the function of the cells expressing these isoforms. For example inhibition of primary or secondary mixed lymphocyte response (MLR) may be determined, such as an in vitro assay or a bioassay for determining the inhibition of primary or secondary MLR in the presence and in the absence of a CD45RO/RB binding molecule and determining the differences in primary MLR inhibition. 
     Alternatively, the in vitro functional modulatory effects can also be determined by measuring the PBMC or T cells or CD4 +  T cells proliferation, production of cytokines, change in the expression of cell surface molecules e.g. following cell activation in MLR, or following stimulation with specific antigen such as tetanus toxoid or other antigens, or with polyclonal stimulators such as phytohemagglutinin (PHA) or anti-CD3 and anti-CD28 antibodies or phorbol esters and Ca 2+  ionophores. The cultures are set up in a similar manner as described for MLR except that instead of allogeneic cells as stimulators soluble antigen or polyclonal stimulators such as those mentioned above are used. T cell proliferation is measured preferably as described above by  3 H-thymidine incorporation. 
     Cytokine production is measured preferably by sandwich ELISA where a cytokine capture antibody is coated on the surface of a 96-well plate, the supernatants from the cultures are added and incubated for 1 hr at room temperature and a detecting antibody specific for the particular cytokine is then added, following a second-step antibody conjugated to an enzyme such as Horseradish peroxidase followed by the corresponding substrate and the absorbance is measured in a plate reader. The change in cell surface molecules may be preferably measured by direct or indirect immunofluorescence after staining the target cells with antibodies specific for a particular cell surface molecule. The antibody can be either directly labeled with fluorochrome or a fluorescently labeled second step antibody specific for the first antibody can be used, and the cells are analysed with a cytofluorimeter. 
     The binding molecule of the invention has a binding specificity for both CD45RO and CD45RB (“CD45RB/RO binding molecule”). 
     Preferably the binding molecule binds to CD45RO isoforms with a dissociation constant (Kd)&lt;20 nM, preferably with a Kd &lt;15 nM or &lt;10 nM, more preferably with a Kd &lt;5 nM. Preferably the binding molecule binds to CD45RB isoforms with a Kd &lt;50 nM, preferably with a Kd &lt;15 nM or &lt;10 nM, more preferably with a Kd &lt;5 nM. 
     In a further preferred embodiment the binding molecule of the invention binds those CD45 isoforms which 
     1) include the A and B epitopes but not the C epitope of the CD45 molecule; and/or
 
2) include the B epitope but not the A and not the C epitope of the CD45 molecule; and/or
 
3) do not include any of the A, B or C epitopes of the CD45 molecule.
 
     In yet a further preferred embodiment the binding molecule of the invention does not bind CD45 isoforms which include 
     1) all of the A, B and C epitopes of the CD45 molecule; and/or
 
2) both the B and C epitopes but not the A epitope of the CD45 molecule.
 
     In further preferred embodiments the binding molecule of the invention further 
     1) recognises memory and in vivo alloactivated T cells; and/or
 
2) binds to its target on human T cells, such as for example PEER cells; wherein said binding preferably is with a Kd &lt;15 nM, more preferably with a Kd &lt;10 nM, most preferably with a Kd &lt;5 nM; and/or
 
3) inhibits in vitro alloreactive T cell function, preferably with an IC 50  of about less than 100 nM, preferably less than 50 nM or 30 nM, more preferably with an IC 50  of about 10 or 5 nM, most preferably with an IC 50  of about 0.5 nM or even 0.1 nM; and/or
 
4) induces cell death through apoptosis in human T lymphocytes; and/or
 
5) induces alloantigen-specific T cell tolerance in vitro; and/or
 
6) prevents lethal xenogeneic graft versus host disease (GvHD) induced in SCID mice by injection of human PBMC when administered in an effective amount; and/or
 
7) binds to T lymphocytes, monocytes, stem cells, natural killer cells and/or granulocytes, but not to platelets or B lymphocytes; and/or
 
8) supports the differentiation of T cells with a characteristic T regulatory cell (Treg) phenotype; and/or
 
9) induces T regulatory cells capable of suppressing naïve T cell activation; and/or
 
10) suppresses the inflammatory process that mediates human allograft skin rejection, in particular, suppresses the inflammatory process that mediates human allograft skin rejection in vivo in SCID mice transplanted with human skin and engrafted with mononuclear splenocytes.
 
     In a further preferred embodiment the binding molecule of the invention binds to the same epitope as the monoclonal antibody “A6” as described by Aversa et al., Cellular Immunology 158, 314-328 (1994). 
     Due to the above-described binding properties and biological activities, such binding molecules of the invention are particularly useful in medicine, for therapy and/or prophylaxis. Diseases in which binding molecules of the invention are particularly useful include autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies, as will be further set out below. 
     We have found that a molecule comprising a polypeptide of SEQ ID NO: 1 and a polypeptide of SEQ ID NO: 2 is a CD45RO/RB binding molecule. We also have found the hypervariable regions CDR1′, CDR2′ and CDR3′ in a CD45RO/RB binding molecule of SEQ ID NO:1, CDR1′ having the amino acid sequence Arg-Ala-Ser-Gln-Asn-Ile-Gly-Thr-Ser-Ile-Gln (RASQNIGTSIQ), CDR2′ having the amino acid sequence Ser-Ser-Ser-Glu-Ser-Ile-Ser (SSSESIS) and CDR3′ having the amino acid sequence Gln-Gln-Ser-Asn-Thr-Trp-Pro-Phe-Thr (QQSNTWPFT). 
     We also have found the hypervariable regions CDR1, CDR2 and CDR3 in a CD45RO/RB binding molecule of SEQ ID NO:2, CDR1 having the amino acid sequence Asn-Tyr-Ile-Ile-His (NYIIH), CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His-Gly-Thr-Lys-Tyr-Asn-Glu-Lys-Phe-Lys-Gly (YFNPYNHGTKYNEKFKG) and CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-Ala-Trp-Phe-Asp-Thr (SGPYAWFDT). 
     CDRs are 3 specific complementary determining regions which are also called hypervariable regions which essentially determine the antigen binding characteristics. These CDRs are part of the variable region, e.g. of SEQ ID NO: 1 or SEQ ID NO: 2, respectively, wherein these CDRs alternate with framework regions (FR&#39;s) e.g. constant regions. A SEQ ID NO: 1 is part of a light chain, e.g. of SEQ ID NO: 3, and a SEQ ID NO:2 is part of a heavy chain, e.g. of SEQ ID NO: 4, in a chimeric antibody according to the present invention. The CDRs of a heavy chain together with the CDRs of an associated light chain essentially constitute the antigen binding site of a molecule of the present invention. It is known that the contribution made by a light chain variable region to the energetics of binding is small compared to that made by the associated heavy chain variable region and that isolated heavy chain variable regions have an antigen binding activity on their own. Such molecules are commonly referred to as single domain antibodies. 
     In one aspect the present invention provides a molecule comprising at least one antigen binding site, e.g. a CD45RO/RB binding molecule, comprising in sequence the hypervariable regions CDR1, CDR2 and CDR3, said CDR1 having the amino acid sequence Asn-Tyr-Ile-Ile-His (NYIIH), said CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His-Gly-Thr-Lys-Tyr-Asn-Glu-Lys-Phe-Lys-Gly (YFNPYNHGTKYNEKFKG) and said CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-Ala-Trp-Phe-Asp-Thr (SGPYAWFDT); e.g. and direct equivalents thereof. 
     In another aspect the present invention provides a molecule comprising at least one antigen binding site, e.g. a CD45RO/RB binding molecule, comprising
     a) a first domain comprising in sequence the hypervariable regions CDR1, CDR2 and CDR3, said CDR1 having the amino acid sequence Asn-Tyr-Ile-Ile-His (NYIIH), said CDR2 having the amino acid sequence Tyr-Phe-Asn-Pro-Tyr-Asn-His-Gly-Thr-Lys-Tyr-Asn-Glu-Lys-Phe-Lys-Gly (YFNPYNHGTKYNEKFKG) and said CDR3 having the amino acid sequence Ser-Gly-Pro-Tyr-Ala-Trp-Phe-Asp-Thr (SGPYAWFDT); and   b) a second domain comprising in sequence the hypervariable regions CDR1′, CDR2′ and CDR3′, CDR1′ having the amino acid sequence Arg-Ala-Ser-Gln-Asn-Ile-Gly-Thr-Ser-Ile-Gln (RASQNIGTSIQ), CDR2′ having the amino acid sequence Ser-Ser-Ser-Glu-Ser-Ile-Ser (SSSESIS) and CDR3′ having the amino acid sequence Gln-Gln-Ser-Asn-Thr-Trp-Pro-Phe-Thr (QQSNTWPFT),
 
e.g. and direct equivalents thereof.
   

     In a preferred embodiment the first domain comprising in sequence the hypervariable regions CDR1, CDR2 and CDR3 is an immunoglobulin heavy chain, and the second domain comprising in sequence the hypervariable regions CDR1′, CDR2′ and CDR3′ is an immunoglobulin light chain. 
     In another aspect the present invention provides a molecule, e.g. a CD45RO/RB binding molecule, comprising a polypeptide of SEQ ID NO: 1 and/or a polypeptide of SEQ ID NO: 2, preferably comprising in one domain a polypeptide of SEQ ID NO: 1 and in another domain a polypeptide of SEQ ID NO: 2, e.g. a chimeric monoclonal antibody, and in another aspect A molecule, e.g. a CD45RO/RB binding molecule, comprising a polypeptide of SEQ ID NO: 3 and/or a polypeptide of SEQ ID NO: 4, preferably comprising in one domain a polypeptide of SEQ ID NO: 3 and in another domain a polypeptide of SEQ ID NO: 4, e.g. a chimeric monoclonal antibody. 
     When the antigen binding site comprises both the first and second domains or a polypeptide of SEQ ID NO: 1 or SEQ ID NO:3, respectively, and a polypeptide of SEQ ID NO: 2 or of SEQ ID NO:4, respectively, these may be located on the same polypeptide, or, preferably each domain may be on a different chain, e.g. the first domain being part of an heavy chain, e.g. immunoglobulin heavy chain, or fragment thereof and the second domain being part of a light chain, e.g. an immunoglobulin light chain or fragment thereof. 
     We have further found that a CD45RO/RB binding molecule according to the present invention is a CD45RO/RB binding molecule in mammalian, e.g. human, body environment. A CD45RO/RB binding molecule according to the present invention can thus be designated as a monoclonal antibody (mAb), wherein the binding activity is determined mainly by the CDR regions as described above, e.g. said CDR regions being associated with other molecules without binding specificity, such as framework, e.g. constant regions, which are substantially of human origin. 
     In another aspect the present invention provides a CD45RO/RB binding molecule which is not the monoclonal antibody “A6” as described by Aversa et al., Cellular Immunology 158, 314-328 (1994), which is incorporated by reference for the passages characterizing A6. 
     In another aspect the present invention provides a CD45RO/RB binding molecule according to the present invention which is a chimeric, a humanised or a fully human monoclonal antibody. 
     Examples of a CD45RO/RB binding molecules include chimeric or humanised antibodies e.g. derived from antibodies as produced by B-cells or hybridomas and or any fragment thereof, e.g. F(ab′) 2  and Fab fragments, as well as single chain or single domain antibodies. A single chain antibody consists of the variable regions of antibody heavy and light chains covalently bound by a peptide linker, usually consisting of from 10 to 30 amino acids, preferably from 15 to 25 amino acids. Therefore, such a structure does not include the constant part of the heavy and light chains and it is believed that the small peptide spacer should be less antigenic than a whole constant part. By a chimeric antibody is meant an antibody in which the constant regions of heavy and light chains or both are of human origin while the variable domains of both heavy and light chains are of non-human (e.g. murine) origin. By a humanised antibody is meant an antibody in which the hypervariable regions (CDRs) are of non-human (e.g. murine) origin while all or substantially all the e.g. the constant regions and the highly conserved parts of the variable regions are of human origins. A humanised antibody may however retain a few amino acids of the murine sequence in the parts of the variable regions adjacent to the hypervariable regions. 
     Hypervariable regions, i.e. CDR&#39;s according to the present invention may be associated with any kind of framework regions, e.g. constant parts of the light and heavy chains, of human origin. Suitable framework regions are e.g. described in “Sequences of proteins of immunological interest”, Kabat, E. A. et al, US department of health and human services, Public health service, National Institute of health. Preferably the constant part of a human heavy chain may be of the IgG1 type, including subtypes, preferably the constant part of a human light chain may be of the κ or λ type, more preferably of the κ type. A preferred constant part of a heavy chain is a polypeptide of SEQ ID NO: 4 (without the CDR1′, CDR2′ and CDR3′ sequence parts which are specified above) and a preferred constant part of a light chain is a polypeptide of SEQ ID NO: 3 (without the CDR1, CDR2 and CDR3 sequence parts which are specified above). 
     We also have found a humanised antibody comprising a light chain variable region of amino acid SEQ ID NO:7 or of amino acid SEQ ID NO:8, which comprises CDR1′, CDR2′ and CDR3′ according to the present invention and a heavy chain variable region of SEQ:ID NO:9 or of SEQ:ID NO:10, which comprises CDR1, CDR2 and CDR3 according to the present invention. 
     In another aspect the present invention provides a humanised antibody comprising a polypeptide of SEQ ID NO:9 or of SEQ ID NO:10 and a polypeptide of SEQ ID NO:7 or of SEQ ID NO:8. 
     In another aspect the present invention provides a humanised antibody comprising
         a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:7,   a polypeptide of SEQ ID NO:9 and a polypeptide of SEQ ID NO:8,   a polypeptide of SEQ ID NO:10 and a polypeptide of SEQ ID NO:7, or   a polypeptide of SEQ ID NO:10 and a polypeptide of SEQ ID NO:8.       

     A polypeptide according to the present invention, e.g. of a herein specified sequence, e.g. of CDR1, CDR2, CDR3, CDR1′, CDR2′CDR3′, or of a SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10 includes direct equivalents of said (poly)peptide (sequence); e.g. including a functional derivative of said polypeptide. Said functional derivative may include covalent modifications of a specified sequence, and/or said functional derivative may include amino acid sequence variants of a specified sequence. 
     “Polypeptide”, if not otherwise specified herein, includes any peptide or protein comprising amino acids joined to each other by peptide bonds, having an amino acid sequence starting at the N-terminal extremity and ending at the C-terminal extremity. Preferably the polypeptide of the present invention is a monoclonal antibody, more preferred is a chimeric (V-grafted) or humanised (CDR-grafted) monoclonal antibody. The humanised (CDR-grafted) monoclonal antibody may or may not include further mutations introduced into the framework (FR) sequences of the acceptor antibody. 
     A functional derivative of a polypeptide as used herein includes a molecule having a qualitative biological activity in common with a polypeptide to the present invention, i.e. having the ability to bind to CD45RO and CD45RB. A functional derivative includes fragments and peptide analogs of a polypeptide according to the present invention. Fragments comprise regions within the sequence of a polypeptide according to the present invention, e.g. of a specified sequence. The term “derivative” is used to define amino acid sequence variants, and covalent modifications of a polypeptide according to the present invention. e.g. of a specified sequence. The functional derivatives of a polypeptide according to the present invention, e.g. of a specified sequence, preferably have at least about 65%, more preferably at least about 75%, even more preferably at least about 85%, most preferably at least about 95% overall sequence homology with the amino acid sequence of a polypeptide according to the present invention, e.g. of a specified sequence, and substantially retain the ability to bind to CD45RO and CD45RB. 
     The term “covalent modification” includes modifications of a polypeptide according to the present invention, e.g. of a specified sequence; or a fragment thereof with an organic proteinaceous or non-proteinaceous derivatizing agent, fusions to heterologous polypeptide sequences, and post-translational modifications. Covalent modified polypeptides, e.g. of a specified sequence, still have the ability bind to CD45RO and CD45RB by crosslinking. Covalent modifications are traditionally introduced by reacting targeted amino acid residues with an organic derivatizing agent that is capable of reacting with selected sides or terminal residues, or by harnessing mechanisms of post-translational modifications that function in selected recombinant host cells. Certain post-translational modifications are the result of the action of recombinant host cells on the expressed polypeptide. Glutaminyl and asparaginyl residues are frequently post-translationally deamidated to the corresponding glutamyl and aspartyl residues. Alternatively, these residues are deaminated under mildly acidic conditions. Other post-translational modifications include hydroxylation of proline and lysine, phosphorylation of hydroxyl groups of seryl, tyrosine or threonyl residues, methylation of the α-amino groups of lysine, arginine, and histidine side chains, see e.g. T. E. Creighton, Proteins: Structure and Molecular Properties, W. H. Freeman &amp; Co., San Francisco, pp. 79-86 (1983). Covalent modifications e.g. include fusion proteins comprising a polypeptide according to the present invention, e.g. of a specified sequence and their amino acid sequence variants, such as immunoadhesins, and N-terminal fusions to heterologous signal sequences. 
     “Homology” with respect to a native polypeptide and its functional derivative is defined herein as the percentage of amino acid residues in the candidate sequence that are identical with the residues of a corresponding native polypeptide, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology, and not considering any conservative substitutions as part of the sequence identity. Neither N- or C-terminal extensions nor insertions shall be construed as reducing identity or homology. Methods and computer programs for the alignment are well known. 
     “Amino acid(s)” refer to all naturally occurring L-α-amino acids, e.g. and including D-amino acids. The amino acids are identified by either the well known single-letter or three-letter designations. 
     The term “amino acid sequence variant” refers to molecules with some differences in their amino acid sequences as compared to a polypeptide according to the present invention, e.g. of a specified sequence. Amino acid sequence variants of a polypeptide according to the present invention, e.g. of a specified sequence, still have the ability to bind to CD45RO and CD45RB. Substitutional variants are those that have at least one amino acid residue removed and a different amino acid inserted in its place at the same position in a polypeptide according to the present invention, e.g. of a specified sequence. These substitutions may be single, where only one amino acid in the molecule has been substituted, or they may be multiple, where two or more amino acids have been substituted in the same molecule. Insertional variants are those with one or more amino acids inserted immediately adjacent to an amino acid at a particular position in a polypeptide according to the present invention, e.g. of a specified sequence. Immediately adjacent to an amino acid means connected to either the α-carboxy or α-amino functional group of the amino acid. Deletional variants are those with one or more amino acids in a polypeptide according to the present invention, e.g. of a specified sequence, removed. Ordinarily, deletional variants will have one or two amino acids deleted in a particular region of the molecule. 
     We also have found the polynucleotide sequences of
         GGCCAGTCAGAACATTGGCACAAGCATACAGTG, encoding the amino acid sequence of CDR1,   TTCTTCTGAGTCTATCTCTGG; encoding the amino acid sequence of CDR 2,   ACAAAGTAATACCTGGCCATTCACGTT encoding the amino acid sequence of CDR 3,   TTATATTATCCACTG, encoding the amino acid sequence of CDR1′,   TTTTAATCCTTACAATCATGGTACTAAGTACAATGAGAAGTTCAAAGGCAG encoding the amino acid sequence of CDR2′,   AGGACCCTATGCCTGGTTTGACACCTG encoding the amino acid sequence of CDR3′,   SEQ ID NO:5 encoding a polypeptide of SEQ ID NO: 1, i.e. the variable region of a light chain of an mAb according to the present invention;   SEQ ID NO:6 encoding a polypeptide of SEQ ID NO:2, i.e. the variable region of the heavy chain of an mAb according to the present invention;   SEQ ID NO:11 encoding a polypeptide of SEQ ID NO:9 i.e. a heavy chain variable region including CDR1, CDR2 and CDR3 according to the present invention;   SEQ ID NO:12 encoding a polypeptide of SEQ ID NO:10, i.e. a heavy chain variable region including CDR1, CDR2 and CDR3 according to the present invention;   SEQ ID NO:13 encoding a polypeptide of SEQ ID NO:7, i.e. a light chain variable region including CDR1′, CDR2′ and CDR3′ according to the present invention; and   SEQ ID NO:14 encoding a polypeptide of SEQ ID NO:8, i.e. a light chain variable region including CDR1′, CDR2′ and CDR3′ according to the present invention.       

     In another aspect the present invention provides isolated polynucleotides comprising polynucleotides encoding a CD45RO/RB binding molecule, e.g. encoding the amino acid sequence of CDR1, CDR2 and CDR3 according to the present invention and/or, preferably and, polynucletides encoding the amino acid sequence of CDR1′, CDR2′ and CDR3′ according to the present invention; and 
     Polynucleotides comprising a polynucleotide of SEQ ID NO: 5 and/or, preferably and, a polynucleotide of SEQ ID NO: 6; and 
     Polynucleotides comprising polynucleotides encoding a polypeptide of SEQ ID NO:7 or SEQ ID NO:8 and a polypeptide of SEQ ID NO:9 or SEQ ID NO:10; e.g. encoding
         a polypeptide of SEQ ID NO:7 and a polypeptide of SEQ ID NO:9,   a polypeptide of SEQ ID NO:7 and a polypeptide of SEQ ID NO:10,   a polypeptide of SEQ ID NO:8 and a polypeptide of SEQ ID NO:9, or   a polypeptide of SEQ ID NO:8 and a polypeptide of SEQ ID NO:10; and       

     Polynucleotides comprising a polynucleotide of SEQ ID NO:11 or of SEQ ID NO:12 and a polynucleotide of SEQ ID NO:13 or a polynucleotide of SEQ ID NO:14, preferably comprising
         a polynucleotide of SEQ ID NO:11 and a polynucleotide of SEQ ID NO:13,   a polynucleotide of SEQ ID NO:11 and a polynucleotide of SEQ ID NO:14,   a polynucleotide of SEQ ID NO:12 and a polynucleotide of SEQ ID NO:13, or   a polynucleotide of SEQ ID NO:12 and a polynucleotide of SEQ ID NO:14.       

     “Polynucleotide”, if not otherwise specified herein, includes any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA, or modified RNA or DNA, including without limitation single and double stranded RNA, and RNA that is a mixture of single- and double-stranded regions. 
     A polynucleotide according to the present invention, e.g. a polynucleotide encoding the amino acid sequence CDR1, CDR2, CDR3, CDR1′, CDR2′, CDR3′, or of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, such as a polynucleotide of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13 or SEQ ID NO:14, respectively, includes allelic variants thereof and/or their complements; e.g. including a polynucleotide that hybridizes to the nucleotide sequence of SEQ ID NO: 5, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13 or SEQ ID NO:14, respectively; e.g. encoding a polypeptide having at least 80% identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, e.g. including a functional derivative of said polypeptide, e.g. said functional derivative having at least 65% homology with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, e.g. said functional derivative including covalent modifications of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, e.g. said functional derivative including amino acid sequence variants of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively; e.g. a SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13 or SEQ ID NO:14, respectively includes a sequence, which as a result of the redundancy (degeneracy) of the genetic code, also encodes a polypeptide of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively, or encodes a polypeptide with an amino acid sequence which has at least 80% identity with the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, respectively. 
     A CD45RO/RB binding molecule, e.g. which is a chimeric or humanised antibody, may be produced by recombinant DNA techniques. Thus, one or more DNA molecules encoding the CD45RO/RB may be constructed, placed under appropriate control sequences and transferred into a suitable host (organism) for expression by an appropriate vector. 
     In another, aspect the present invention provides a polynucleotide which encodes a single, heavy and/or a light chain of a CD45RO/RB binding molecule according to the present invention; and the use of a polynucleotide according to the present invention for the production of a CD45RO/RB binding molecule according to the present invention by recombinant means. 
     A CD45RO/RB binding molecule may be obtained according, e.g. analogously, to a method as conventional together with the information provided herein, e.g. with the knowledge of the amino acid sequence of the hypervariable or variable regions and the polynucleotide sequences encoding these regions. A method for constructing a variable domain gene is e.g. described in EP 239 400 and may be briefly summarized as follows: A gene encoding a variable region of a mAb of whatever specificity may be cloned. The DNA segments encoding the framework and hypervariable regions are determined and the DNA segments encoding the hypervariable regions are removed. Double stranded synthetic CDR cassettes are prepared by DNA synthesis according to the CDR and CDR′ sequences as specified herein. These cassettes are provided with sticky ends so that they can be ligated at junctions of a desired framework of human origin. Polynucleotides encoding single chain antibodies may also be prepared according to, e.g. analogously, to a method as conventional. A polynucleotide according to the present invention thus prepared may be conveniently transferred into an appropriate expression vector. 
     Appropriate cell lines may be found according, e.g. analogously, to a method as conventional. Expression vectors, e.g. comprising suitable promotor(s) and genes encoding heavy and light chain constant parts are known e.g. and are commercially available. Appropriate hosts are known or may be found according, e.g. analogously, to a method as conventional and include cell culture or transgenic animals. 
     In another aspect the present invention provides an expression vector comprising a polynucleotide encoding a CD45RO/RB binding molecule according to the present invention, e.g. of sequence SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17 or SEQ ID NO:18. 
     In another aspect the present invention provides
         An expression system comprising a polynucleotide according to the present invention wherein said expression system or part thereof is capable of producing a CD45RO/RB binding molecule according to the present invention, when said expression system or part thereof is present in a compatible host cell;
 
and
   An isolated host cell which comprises an expression system as defined above.       

     We have further found that a CD45RO/RB binding molecule according to the present invention inhibit primary alloimmune responses in a dose-dependent fashion as determined by in vitro MLR. The results indicate that the cells which had been alloactivated in the presence of a CD45RO/RB binding molecule according to the present invention are impaired in their responses to alloantigen. This confirms the indication that a CD45RO/RB binding molecule according to the present invention can act directly on the effector alloreactive T cells and modulate their function. In addition, the functional properties of T cells derived from the primary MLR were further studied in restimulation experiments in secondary MLR, using specific stimulator cells or third-party stimulators to assess the specificity of the observed functional effects. We have found that the cells derived from primary MLRs in which a CD45RO/RB binding molecule according to the present invention is present, were impaired in their ability to respond to subsequent optimal stimulation with specific stimulator cells, although there was no antibody added to the secondary cultures. The specificity of the inhibition was demonstrated by the ability of cells treated with a CD45RO/RB binding molecule according to the present invention to respond normally to stimulator cells from unrelated third-party donors. Restimulation experiments using T cells derived from primary MLR cultures thus indicate that the cells which had been alloactivated a CD45RO/RB binding molecule according to the present invention are hyporesponsive, i.e. tolerant, to the original alloantigen. Further biological activities are described in examples 7, and 9 to 12. 
     Furthermore we have found that cell proliferation in cells pre-treated with a CD45RO/RB binding molecule according to the present invention could be rescued by exogenous IL-2. This indicates that treatment of alloreactive T cells with a CD45RO/RB binding molecule according to the present invention induces a state of tolerance. Indeed, the reduced proliferative responses observed in cells treated with a CD45RO/RB binding molecule according to the present invention, was due to impairement of T cell function, and these cells were able to respond to exogenous IL-2, indicating that these cells are in an anergic, true unresponsive state. The specificity of this response was shown by the ability of cells treated with a CD45RO/RB binding molecule according to the present invention to proliferate normally to unrelated donor cells to the level of the control treated cells. 
     In addition experiments indicate that the binding of a CD45RO/RB binding molecule according to the present invention to CD45RO and CD45RB may inhibit the memory responses of peripheral blood mononuclear cells (PBMC) from immunized donors to specific recall antigen. Binding of a CD45RO/RB binding molecule according to the present invention to CD45RO and CD45RB thus is also effective in inhibiting memory responses to soluble Ag. The ability of a CD45RO/RB binding molecule according to the present invention to inhibit recall responses to tetanus in PBMC from immunized donors indicate that a CD45RO/RB binding molecule according to the present invention is able to target and modulate the activation of memory T cells. E.g. these data indicate that a CD45RO/RB binding molecule according to the present invention in addition to recognizing alloreactive and activated T cells is able to modulate their function, resulting in induction of T cell anergy. This property may be important in treatment of ongoing immune responses to autoantigens and allergens and possibly to alloantigens as seen in autoimmune diseases, allergy and chronic rejection, and diseases, such as psoriasis, inflammatory bowel disease, where memory responses play a role in the maintenance of disease state. It is believed to be an important feature in a disease situation, such as in autoimmune diseases in which memory responses to autoantigens may play a major role for the disease maintenance. 
     We have also found that a CD45RO/RB binding molecule according to the present invention may modulate T cell proliferative responses in a mixed lymphocyte response (MLR) in vivo, i.e. a CD45RO/RB binding molecule according to the present invention was found to have corresponding inhibitory properties in vivo testing. 
     A CD45RO/RB binding molecule according to the present invention may thus have immunosuppressive and tolerogenic properties and may be useful for in vivo and ex-vivo tolerance induction to alloantigens, autoantigens, allergens and bacterial flora antigens, e.g. a CD45RO/RB binding molecule according to the present invention may be useful in the treatment and prophylaxis of diseases e.g. including autoimmune diseases, such as, but not limited to, rheumatoid arthritis, autoimmune thyroditis, Graves disease, type I and type II diabetes, multiple sclerosis, systemic lupus erythematosus, Sjögren syndrome, scleroderma, autoimmune gastritis, glomerulonephritis, transplant rejection, e.g. organ and tissue allograft and xenograft rejection, graft versus host disease (GVHD), and also psoriasis, inflammatory bowel disease and allergies. 
     In another aspect the present invention provides the use of a CD45RO/RB binding molecule according to the present invention as a pharmaceutical, e.g. in the treatment and prophylaxis of autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies. 
     In another aspect the present invention provides a CD45RO/RB binding molecule according to the present invention for the production of a medicament in the treatment and prophylaxis of diseases associated with autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies. 
     In another aspect the present invention provides a pharmaceutical composition comprising a CD45RO/RB binding molecule according to the present invention in association with at least one pharmaceutically acceptable carrier or diluent. 
     A pharmaceutical composition may comprise further, e.g. active, ingredients, e.g. other immunomodulatory antibodies such as, but not confined to anti-ICOS, anti-CD154, anti-CD134L or recombinant proteins such as, but not confined to rCTLA-4 (CD152), rOX40 (CD134), or immunomodulatory compounds such as, but not confined to cyclosporin A, FTY720, RAD, rapamycin, FK506, 15-deoxyspergualin, steroids. 
     In another aspect the present invention provides a method of treatment and/or prophylaxis of diseases associated with autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies comprising administering to a subject in need of such treatment and/or prophylaxis an effective amount of a CD45RO/RB binding molecule according to the present invention, e.g. in the form of a pharmaceutical composition according to the present invention. 
     Autoimune diseases to be treated with binding molecule of the present invention further include, but are not limited to, rheumatoid arthritis, autoimmune thyroditis, Graves disease, type I and type II diabetes, multiple sclerosis, systemic lupus erythematosus, Sjögren syndrome, scleroderma, autoimmune gastritis, glomerulonephritis; transplant rejection, e.g. organ and tissue allograft and xenograft rejection and graft-versus-host disease (GVHD). 
     EXAMPLES 
     The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention. In the following examples all temperatures are in degree Celsius. 
     The “candidate mAb” or “chimeric antibody” is a CD45RO/RB binding molecule according to the present invention comprising light chain of SEQ ID NO:3 and heavy chain of SEQ ID NO:4. 
     The “humanised antibody” is a CD45RO/RB binding molecule according to the present invention comprising a polypeptide of SEQ ID NO:8 and polypeptide of SEQ ID NO:9 or a polypeptide of SEQ ID NO:8 and a polypeptide of SEQ ID NO:10. 
     The following abbreviations are used: 
     APC antigen presenting cell
 
c.p.m. counts per minute
 
ELISA enzyme linked immuno-sorbant assay
 
FACS fluorescence activated cell sorting
 
Fc fragment crystallizable
 
F(ab′) 2  fragment antigen-binding; bivalent
 
FITC fluorescein isothiocyanate
 
FBS foetal bovine serum
 
GVHD graft-vs-host disease
 
HCMV human cytomegalovirus promoter
 
IFN-γ interferon gamma
 
IgE immunoglobulin isotype E
 
IgG immunoglobulin isotype G
 
IL-2 interleukin-2
 
IU units
 
MLR mixed lymphocyte reaction
 
MLC mixed lymphocyte culture
 
MP1 matrix protein 1 from  hemophilus influenza  
 
PBS phosphate-buffered saline
 
PBL peripheral blood leukocytes
 
PBMC peripheral blood mononuclear cells
 
PCR polymerase chain reaction
 
SCID severe combined immunodeficiency
 
T reg  T regulatory cells
 
xGVHD xeno-graft-vs-host disease
 
     Example 1 
     Primary Mixed Lymphocyte Response (MLR) 
     Cells 
     Blood samples are obtained from healthy human donors. Peripheral blood mononuclear cells (PBMC) are isolated by centrifugation over Ficoll-Hypaque (Pharmacia LKB) from leukocytes from whole peripheral blood, leukopheresis or buffy coats with known blood type, but unknown HLA type. In some MLR experiments, PBMC are directly used as the stimulator cells after the irradiation at 40 Gy. In the other experiments, T cells were depleted from PBMC by using CD2 or CD3 Dynabeads (Dynal, Oslo, Norway). Beads and contaminating cells are removed by magnetic field. T cell-depleted PBMC are used as simulator cells after the irradiation. 
     PBMC, CD3 +  T cells or CD4 +  T cells are used as the responder cells in MLR. Cells are prepared from different donors to stimulator cells. CD3 +  T cells are purified by negative selection using anti-CD16 mAb (Zymed, Calif.), goat anti-mouse IgG Dynabeads, anti-CD14 Dynabeads, CD19 Dynabeads. In addition anti-CD8 Dynabeads are used to purify CD4 +  T cells. The cells obtained are analyzed by FACScan or FACSCalibur (Becton Dickinson &amp; Co., CA) and the purity of the cells obtained was &gt;75%. Cells are suspended in RPMI1640 medium, supplemented with 10% heat-inactivated FBS, penicillin, streptomycin and L-glutamine. 
     Reagents 
     The chimeric anti-CD45R0/RB mAb “candidate mAb” and an isotype matched control chimeric antibody is also generated. Mouse (Human) control IgG 1  antibody specific for KLH (keyhole limpet hemocyanin) or recombinant human IL-10 is purchased from BD Pharmingen (San Diego, Calif.). Anti-human CD154 mAb 5c8 is according to Lederman et al 1992. 
     Primary Mixed Lymphocyte Response (MLR) 
     Aliquots of 1×10 5  PBMC or 5×10 4  of CD3 +  or CD4 +  cells are mixed with 1×10 5  irradiated PBMC or 5×10 4  T cells-depleted irradiated (50 Gy) PBMC in the each well of 96-well culture plates (Costar, Cambridge, Mass.) in the presence of the indicated mAb or absence of Ab. In some experiments, F(ab′) 2  fragment of goat anti-mouse Ig or goat anti-human Ig specific for Fc portion (Jackson ImmunoResearch, West Grove, Pa.) is added at 10 μg/ml in addition to the candidate mAb To ensure optimal in vitro cross-linking of the target CD45 molecules. The mixed cells are cultured for 4 or 5 days at 37° C. in 5% CO 2  and proliferation is determined by pulsing the cells with  3 H-thymidine for the last 16-20 hours of culture. Other experiments are similar to those described above, but with the following exceptions: 1) Medium used is EX-VIVO (Bio-Whittaker) containing 10% FBS and 1% human plasma; 2) Anti-mouse total IgG (5 μg/ml) is used as secondary cross-linking step; 3) Irradiation of stimulator cells is 60 Gy. 
     Primary MLR is performed in the presence of the “candidate mAb” or control chimeric IgG 1  (10 μg/ml) both with a second step reagent, F(ab′) 2  fragment of goat anti-human Ig specific for Fc portion (10 μg/ml). Percentage inhibition by the “candidate mAb” is calculated in comparison with the cell proliferation in the presence of control IgG 1 . Results are shown in TABLE 1 below: 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Inhibition of primary MLR by 10 μg/ml 
               
               
                 of a candidate mAb according to the present invention 
               
            
           
           
               
               
               
               
            
               
                   
                 Responder 
                 Stimulator (Irr. PBMC) 
                 % of Inhibition 
               
               
                   
                   
               
               
                   
                 #211 CD4 
                 #219 CD3 
                 63.51 
               
               
                   
                 #220 CD4 
                 #219 CD3 depl. 
                 63.07 
               
               
                   
                 #227 CD4 
                 #220 CD3 depl. 
                 65.96 
               
               
                   
                 #229 CD4 
                 #219 CD3 depl. 
                 50.76 
               
            
           
           
               
               
               
            
               
                   
                 Average ± SD 
                 60.83 ± 6.83* 
               
               
                   
                   
               
               
                   
                 *Significantly different from control value (P &lt; 0.001) 
               
            
           
         
       
     
     A candidate mAb according to the present invention inhibits primary MLR as can be seen from TABLE 1. The average inhibitory effect is 60.83±6.83% in four different donors-derived CD4 +  T cells and statistically significant. 
     The inhibition of primary MLR by the “candidate mAb” is shown to be dose-dependent in the range of 0.001 and 10 μg/ml of the “candidate mAb” as shown in  FIG. 1 . 
     The IC 50  for the inhibition of primary MLR by a “candidate mAb” is determined from the results of three separate MLR experiments using one donor PBMC as responder cells. Thus, responder CD4 +  T cells from Donor #229 and #219 and irradiated PBMC depleted of T cells as stimulators are mixed in the presence of a “candidate mAb” or control chimeric Ab with 10 μg/ml of F(ab′) 2  fragment of goat anti-human Ig. Experiments are repeated 3 times and percentage of proliferation in the presence of a “candidate mAb” is calculated in comparison with the T cell proliferation in the presence of control Ab. IC 50  value is determined using Origin (V. 6.0®). The cellular activity IC 50  value is calculated to be 0.87±0.35 nM (0.13±0.052 μg/ml). 
     Example 2 
     Secondary MLR 
     In order to assess whether a “candidate mAb” induces unresponsiveness of CD4 +  T cells to specific alloantigens, secondary MLR is performed in the absence of any antibodies after the primary MLC. CD4 +  T cells are cultured with irradiated allogeneic stimulator cells (T cells-depleted PBMC) in the presence of the indicated antibody in 96-well culture plates for 10 days (primary MLC). Then, cells are collected, layered on a Ficoll-Hypaque gradient to remove dead cells, washed twice with RPMI, and restimulated with the same stimulator, 3 rd  party stimulator cells or IL-2 (50 U/ml). The cells are cultured for 3 days and the proliferative response is determined by pulsing the cells with  3 H-thymidine for the last 16-20 hours of culture. 
     Specifically, CD4 +  T cells are cultured with irradiated allogeneic stimulator cells (T cells-depleted PBMC taken from other donors) in the presence of 10 μg/ml of the “candidate mAb”, control IgG1 chimeric Ab and F(ab′) 2  fragment of goat anti-human Ig. Primary MLR proliferation is determined on day 5. For secondary MLR, the responder and stimulator cells are cultured for 10 days in the presence of the “candidate mAb”, then the cells are harvested, washed twice in RPMI1640 and restimulated with specific stimulator, third-party stimulators or IL-2 (50 U/ml) in the absence of any Ab. Cell proliferation is determined on day 3. Results set out in TABLE 2: 
     
       
         
           
               
               
               
             
               
                   
                 TABLE 2 
               
               
                   
                   
               
               
                   
                 Responder CD4+ T cells Donor # 
                 % Inhibition of 2 ry  MLR 
               
               
                   
                   
               
             
            
               
                   
                 #211 
                 49.90* 
               
               
                   
                 #220 
                 59.33* 
               
               
                   
                 #227 
                 58.68* 
               
               
                   
                   
               
               
                   
                 *Significantly different from control value (p = &lt;0.001 determined by t-test, SigmaStat V.2.03). 
               
               
                   
                 # p = &lt;0.046 
               
            
           
         
       
     
     In order to test whether the impaired proliferation is due to unresponsivess as a consequence of the treatment with a “candidate mAb”, the cells derived from primary MLR are cultured in the presence of IL-2 (50 U/ml). Addition of IL-2 results in the rescue of proliferative responses of the T cells which had been treated with a “candidate mAb” in primary MLR, to levels similar to those observed in the presence of IgG 1  control Ab. These data indicate that the impaired secondary response in T cells treated with a “candidate mAb” is due to to functional alteration of the responder T cells which become unresponsive to the specific stimulator cells. 
     Percentage inhibition is calculated according to the following formula: 
     
       
         
           
             
               
                 
                   
                     c 
                     . 
                     p 
                     . 
                     m 
                     . 
                     
                       
                           
                       
                        
                       
                           
                       
                     
                      
                     with 
                   
                    
                   
                       
                   
                    
                   control 
                    
                   
                       
                   
                    
                   Ab 
                 
                 - 
                 
                   
                     c 
                     . 
                     p 
                     . 
                     m 
                     . 
                     
                         
                     
                      
                     with 
                   
                    
                   
                       
                   
                    
                   
                     “ 
                     
                       candidate 
                        
                       
                           
                       
                        
                       mAb 
                     
                   
                 
               
               
                 
                   c 
                   . 
                   p 
                   . 
                   m 
                   . 
                   
                       
                   
                    
                   with 
                 
                  
                 
                     
                 
                  
                 control 
                  
                 
                     
                 
                  
                 Ab 
               
             
             × 
             100 
           
         
       
     
     Statistical analysis is performed using SigmaStat (Vers. 2.03). 
     The data is analyzed by two-way ANOVA followed by Dunnett method. In all test procedures probabilities &lt;0.05 are considered as significant. In some experiments t-test is used (SigmaStat V. 2.03). 
     Example 3 
     In Vivo Survival Studies in SCID-Mice 
     Engraftment of hu-PBL in SCID Mice 
     Human peripheral blood mononuclear cells (PBMC) are injected intraperitoneally into SCID mice C.B 17/GbmsTac-Prkdc scid  Lyst bg  mice (Taconic, Germantown, N.Y.) in an amount sufficient to induce a lethal xenogeneic graft-versus-host disease (xGvHD) in &gt;90% of the mice within 4 weeks after cell transfer. Such treated SCID mice are hereinafter designated as hu-PBL-SCID mice 
     Mab-Treatment of hu-PBL-SCID Mice 
     Hu-PBL-SCID mice are treated with a “candidate mAb” or mouse or chimeric isotype matched mAb controls at day 0, immediately after PBMC injection, at day 3, day 7 and at weekly intervals thereafter. Mabs are delivered subcutaneously in 100 μl PBS at a final concentration of 5 mg/kg body weight. The treatment was stopped when all control mice were dead. 
     Evaluation of Treatment Results 
     The main criterion to assess the efficacy of a “candidate mAb” in this study was the survival of the hu-PBL-SCID mice. The significance of the results is evaluated by the statistical method of survival analysis using the Log-rank test (Mantel method) with the help of the Systat v9.01 software. The method of survival analysis is a non-parametric test, which not only consider whether a particular mouse is still alive but also whether if it was sacrificed for reasons irrelevant to the treatment/disease such as the requirement of perform in vitro analysis with its organs/cells. Biopsies of liver, lung, kidney and spleen are obtained from dead mice for further evaluation. In addition, hu-PBL-SCID mice are weighed at the beginning (before cell transfer) and throughout (every two days) the experiment as an indirect estimation of their health status. Linear regression lines were generated using the body weight versus days post-PBMC transfer values obtained from each mouse and subsequently, their slopes (control versus anti-CD45 treated mice) were compared using the non-parametric Mann-Whitney test. 
     Results 
     All hu-PBL-SCID mice treated with mouse mAb controls had infiltrated human leukocytes in the lung, liver and spleen and died (4/4) within ca. 2 to 3 weeks after cell transfer. Death is a likely consequence of xGvHD. Control mAb-treated mice furthermore lost weight in a linear manner, ca. 10% and more within 3 weeks. 
     All hu-PBL-SCID mice treated with a “candidate mAb” survived (4/4) without any apparent sign of disease more than 4 weeks, even although “candidate mAb”-treatment was stopped after 3 weeks. “Candidate mAb”-treated mice increased weight in a linear manner, up to ca. 5% within 4 weeks. 
     Example 4 
     Expression of Antibodies of the Invention 
     Expression of Humanised Antibody Comprising a SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10 
     Expression vectors according to the plasmid map shown in  FIGS. 2 to 5  are constructed, comprising the corresponding nucleotides encoding the amino acid sequence of humanised light chain variable region humV1 (SEQ ID NO:7), humanised light chain variable region humV2 (SEQ ID NO:8), humanised heavy chain variable region VHE (SEQ ID NO:9), or humanised heavy chain variable region VHQ (SEQ ID NO:10), respectively. These expression vectors have the DNA (nucleotide) sequences SEQ ID NO 15, SEQ ID NO 16, SEQ ID NO 17, or SEQ ID NO 18, respectively. 
     Construction of Humanised Antibody Heavy and Light Chain Expression Vectors 
     Human Kappa Light Chain Expression Vectors for Versions VLh and VLm 
     In order to construct the final expression vector encoding for the complete humanised light chain of human kappa isotype, DNA fragments encoding the complete light chain variable regions (VLh and VLm) were excised from the VLh and VLm containing PCR-Script cloning vectors (Stratagene) (VLm region) using HindIII and BgIII. The gel-purified fragments were then subcloned into the HindIII and BamHI sites of C21-HCMV Kappa expression vector which was created during construction of the humanised anti-IgE antibody TESC-21 (Kolbinger et al 1993) and which originally received from M. Bendig (MRC Collaborative Centre, London, UK) (Maeda et al. 1991). The ligation products were purified by phenol/chloroform extraction, and electroporated into electrocoporation-competent Epicurian Coli® XL1-Blue strain (Cat. No #200228, Stratagene). After plating on LB/amp agar plates overnight at 37° C., each 12 colonies were picked to prepare plasmid DNA from a 3 ml culture using the BioRobot 9600 (Qiagen). This yielded the light chain expression vectors for the humanised antibody versions VLh and VLm, respectively, as further described in the Figures. 
     Human Gamma-1 Heavy Chain Expression Vectors for VHQ 
     For the construction of the VHQ expression vector, a step-wise approach was taken. First, the complete variable region of VHQ was assembled by PCR according to the methology as described in Kolbinger et al 1993 (Protein Eng. 1993 November; 6(8):971-80) and subcloned into the C21-HCMV-gamma-1 expression from which the C21 insert had been removed using the same enzymes. A HindIII/BamHI fragment of PCRScript clone VHQ containing the complete variable region was then subcloned into expression vector C21-HCMV-gamma-1 cleaved with the same enzymes. This yielded the final expression vector for the humanised antibody version VHQ. 
     Human Gamma-1 Heavy Chain Expression Vectors for VHE 
     The construction of the final VHE expression vector encoding for the complete humanised heavy chain of human gamma-1 isotype was achieved by directly ligating a HindIII and BamHI restricted PCR fragment encoding the variable region into the HindIII and BamHI sites of C21-HCMV gamma-1 expression vector which was created during construction of the humanised anti-IgE antibody TESC-21 (Kolbinger et al 1993) and which was also originally received from M. Bendig (MRC Collaborative Centre, London, UK) (Maeda et al. 1991). 
     Transient Expression in COS Cells 
     The following transfection protocol is adapted for adherent COS cells in 150 mm cell culture dishes, using SuperFect™ Transfection Reagent (Cat. No 301305, Qiagen). The four different expression vectors described above are used for transient transfection of cells. For expression of humanised antibody, each of two clones containing heavy chain inserts (VHE or VHQ, respectively) are co-transfected into cells with each of the two clones encoding for the light chains (humV1 or humV2, respectively), in total 4 different combinations of heavy and light chain expression vectors (VHE/humV1, VHE/humV2, VHQ/humV1 and VHQ/humV2). Before transfection, the plasmids are linearized with the restriction endonuclease Pvul which cleaves in the region encoding the resistance gene for ampicillin. The day before transfection, 4×10 6  COS cells in 30 ml of fresh culture medium are seeded in 150 mm cell culture dishes. Seeding at this cell density generally yielded 80% confluency after 24 hours. On the day of transfection, four different combinations of linearized heavy- and light-chain DNA expression vectors (15 μg each) are diluted in a total volume of 900 μl of fresh medium without serum and antibiotics. 180 μl of SuperFect Transfection Reagent is then mixed thoroughly with the DNA solution. The DNA mixture is incubated for 10 min at room temperature to allow complex formation. While complex formation takes place, the growth medium is removed from COS cell cultures, and cells are washed once with PBS. 9 ml of fresh culture medium (containing 10% FBS and antibiotics) are then added to each reaction tube containing the transfection complexes and well mixed. The final preparation is immediately transferred to each of 4 cultures to be transfected and gently mixed. Cell cultures are then incubated with the DNA complexes for 3 hours at 37° C. and 5% CO2. After incubation, the medium containing transfection complexes is removed and replaced with 30 ml of fresh culture medium. At 48 hr post transfection, the culture supernatants are harvested. 
     Concentration of Culture Supernatants 
     For ELISA and FACS analysis, the culture supernatants collected from COS cells transfected with heavy- and light-chain plasmids are concentrated as follows. 10 ml of each supernatant are added to Centriprep YM-50 Centrifugal Filter Devices (Cat. No 4310, Millipore) as described by the manufacturer. The Centriprep filters are centrifuged for 10 min at 3000 rpm at room temperature. The centrifugation step is then repeated again with the remaining 20 ml of supernatant using only 5 min of centrifugation and supervising the concentration evolution. The intermediate 500 μl of concentrated supernatant is recovered, transferred to new Microcon Centrifugal Filter Devices (Cat. No 42412, Microcon) and further concentrated following the manufacturer&#39;s protocol. The concentrated supernatants are centrifuged four times for 24 min at 3000 rpm at room temperature, one time for 10 min at 6000 rpm and then, three times for 5 min, always supervising the concentration evolution. The final volume of concentrated conditioned medium achieved is 100-120 μl corresponding to a 250 to 300-fold concentration of original culture medium and is stored at 4° C. until use. For comparison and control, culture medium from untransfected cells is similarly concentrated, using the same centrifugation protocol described above. 
     Generation of Stable Sp2/0 Myeloma Transfectants Secreting Humanised Anti-CD45RO/RB Antibodies 
     The mouse myeloma cell line Sp2/0 (ATCC, CRL-1581) is electroporated with vectors encoding heavy (VHE or VHQ) and light (humV1 or humV2) chain of the CD45RO/RB binding humanised antibodies. Four different combinations of heavy and light chain expression vectors (VHE/humV1, VHE/humV2, VHQ/humV1 and VHQ/humV2) are transfected according to the following protocol: 20 μg supercoiled DNA of each plasmid is mixed in an electroporation cuvette (0.4 cm gap) with 8×10 6  live Sp2/0 cells suspended in DMEM/10% FCS culture medium. Electroporation settings are 1500 V, 25 μF using a BioRad GenePulser instrument. After electroporation, cells are cultured for 20 h in culture medium (DMEM supplemented with 10% FCS penicillin, streptomycin and L-glutamine). On day two the selection drug G418 (Cat. No 10131-019, Gibco) is added to a final concentration of 1 mg active drug/ml and the cells are distributed into one 96-well plate, 200 μl each well with approx. 10 5  cells per well. Ten to 15 days later, G418-surviving clones are expanded in G418-containing medium. Secretion of humanised mAbs from these transfectants is assessed by ELISA, using a coating antibody goat anti-human IgG/Fcγ (Cat. No 109-005-098, Jackson Labs) and a peroxidase-coupled antibody against human kappa light chain (Cat. No A-7164, Sigma). Transfectants, which score positive in this assay are selected for a comparison of productivity on a per cell per day basis, again using ELISA (see below). The best clone of each transfectant is selected for immediate subcloning by limiting dilution, using a seeding density of 1 cell per well. Productivity of G418-surviving subclones is again determined as described above. Subclones are expanded in G418-containing selection medium, until the culture volume reaches 150 ml, at which stage the culture is continued without G418 in flasks destined to feed roller bottles. 
     After the first transfection and selection, stable transfectants grow out of the 96-well plates at a frequency of 20.8% for VHE/humV1, 11.5% for VHQ/humV1, 18.8% for VHE/humV2 and 7.3% for VHQ/humV2. After two rounds of subcloning the best two producers are clone 1.33.25 (3.87 pg/cell/day) and clone 1.33.26 (3.43 pg/cell/day) for VHE/humV1 and clone 12.1.4 (1.19 pg/cell/day) and clone 12.1.20 (1.05 pg/cell/day) for VHQ/humV1. The stable Sp2/0 transfectants for VHE/humV1 and VHQ/humV1 are subsequently expanded for antibody production and purification. 
     The antibodies are purified from supernatants of stably transfected SP2/0 myeloma cell lines containing 10% FCS by a combination of affinity chromatography using an immobilized anti-human IgGFc matrix and size-exclusion chromatography. If required, endotoxin is removed using an Acticlean Etox column (Sterogene Bioseparations). 
     Example 5 
     Determination of Recombinant Human IgG Expression by ELISA 
     To determine IgG concentrations of recombinant human antibody expressed in the culture supernatants, a sandwich ELISA protocol has been developed and optimized using human IgG as standard. Flat bottom 96-well microtiter plates (Cat. No 4-39454, Nunc Immunoplate Maxisorp) are coated overnight at 4° C. with 100 μl of goat anti-human IgG (whole molecule, Cat. No 11011, SIGMA) at the final concentration of 0.5 μg/ml in PBS. Wells are then washed 3 times with washing buffer (PBS containing 0.05% Tween 20) and blocked for 1.5 hours at 37° C. with blocking buffer (0.5% BSA in PBS). After 3 washing cycles, the antibody samples and the standard human IgG (Cat. No. 14506, SIGMA) are prepared by serial 1.5-fold dilution in blocking buffer. 100 μl of diluted samples or standard are transfered in duplicate to the coated plate and incubated for 1 hour at room temperature. After incubation, the plates are washed 3 times with washing buffer and subsequently incubated for 1 hour with 100 μl of horseradish peroxidase-conjugated goat anti-human IgG kappa-light chain (Cat. No A-7164, SIGMA) diluted at 1/4000 in blocking buffer. Control wells received 100 μl of blocking buffer or concentrated normal culture medium. After washing, the colorimetric quantification of bound peroxidase in the sample and standard wells is performed, using a TMB Peroxidase EIA Substrate Kit (Cat. No 172-1067, Bio-Rad) according to the manufacturer&#39;s instructions. The peroxidase mixture is added at 100 μl per well and incubated for 30 min at room temperature in the dark. The colorimetric reaction is stopped by addition of 100 μl of 1 M sulfuric acid and the absorbance in each well is read at 450 nm, using an ELISA plate reader (Model 3350-UV, BioRad). 
     With a correlation coefficient of 0.998 for the IgG standard curve, the following concentrations are determined for the four different culture concentrates (ca. 250-300 fold concentrated) obtained from transfected COS cells: 
     VHE/humV1 supernatant=8.26 μg/ml
 
VHE/humV2 supernatant=6.27 μg/ml
 
VHQ/humV1 supernatant=5.3 μg/ml
 
VHQ/humV2 supernatant=5.56 μg/ml
 
     Example 6 
     FACS Competition Analysis (Binding Affinity) 
     The human T-cell line PEER is chosen as the target cell for FACS analysis because it expressed the CD45 antigen on its cell surface. To analyze the binding affinity of humanised antibody supernatants, competition experiments using FITC-labeled chimeric antibody as a reference are performed and compared with the inhibition of purified mouse antibody and of chimeric antibody. PEER cell cultures are centrifuged for 10 seconds at 3000 rpm and the medium is removed. Cells are resuspended in FACS buffer (PBS containing 1% FBS and 0.1% sodium azide) and seeded into 96-well round-bottom microtitter plate at a cell density of 1×10 5  cells per well. The plate is centrifuged and the supernatant is discarded. For blocking studies, 25 μl of concentrated untransfected medium or isotype matched control antibody (negative controls), unlabeled mouse antibody or chimeric antibody (positive controls) as well as concentrated supernatant containing the various combinations of humanised antibody (samples), is first added in each well at the indicated concentrations in the text. After 1 hour of incubation at 4° C., PEER cells are washed with 200 μl of FACS buffer by centrifugation. Cells are subsequently incubated for 1 hour at 4° C. with chimeric antibody conjugated with FITC in 25 μl of FACS buffer at the final concentration of 20 μg/ml. Cells are washed and resuspended in 300 μl of FACS buffer containing 2 μg/ml propidium iodide which allows gating of viable cells. The cell preparations are analyzed on a flow cytometer (FACSCalibur, Becton Dickinson). 
     FACS analyses indicate a dose-dependent blockade of fluorochrome-labeled chimeric antibody by the concentrated humanised antibody culture supernatants. No dose-dependent blockade of chimeric antibody binding is seen with the isotype matched control antibody, indicating that the blocking effect by the different humanised antibody combinations is epitope specific and that epitope specificity appears to be retained after the humanisation process. 
     Undiluted supernatant from the above mentioned SP2/0 transfectants or chimeric antibody (positive controls) or isotype matched control antibody (negative controls) at 2 μg/ml in culture medium are incubated with 1.5×10 5  PEER cells in 100 μl for 30 min at 4° C. Then, 100 μl PBS containing FITC-labeled chimeric antibody is added to each sample and incubation at 4° C. continues for another 30 minutes. After washing, cells are resuspended in FACS-PBS containing 1 μg/ml 7-Amino-Actinomycin D and analyzed by flow cytometry using a Becton Dickinson FACSCalibur instrument and the CellQuest Pro Software. Gating was on live cells, i.e. 7-Amino-Actinomycin D—negative events. 
     FACS analyses show that unlabeled humanised CD45RB/RO binding molecules, e.g. VHE/humV1 and VHQ/humV1 but not the isotype matched control antibody compete with FITC-labeled chimeric antibody for binding to the human CD45-positive T cell line PEER. 
     Example 7 
     Biological Activities of CD45RB/RO Binding Molecules 
     In this study, we have addressed whether CD45RB/RO binding chimeric antibody, when present in cultures of polyclonally activated primary human T cells (i) supports the differentiation of T cells with a characteristic Treg phenotype, (ii) prevents or enhances apoptosis following T cell activation, and (iii) affects expression of subset-specific antigens and receptors after restimulation. 
     CD45RB/RO Binding Chimeric Antibody Enhances Cell Death in Polyclonally Activated T Cells 
     Primary T cells (mixture of CD4+ and CD8+ T subsets) were subjected to activation by anti-CD3 plus anti-CD28 mAb (200 ng/ml each) in the presence or absence (=control) of CD45RB/RO binding chimeric antibody. Excess antibodies were removed by washing on day 2. 7-amino-actinomycin D (7-AAD) as a DNA-staining dye taken up by apoptotic and necrotic cells was used to measure cell death following activation. The results show that activation of T cells in the presence of CD45RB/RO binding chimeric antibody increased the fraction of 7-AAD positive cells than two-fold on day 2 after activation. On day 7, the portion of 7-AAD positive cells was again similar in CD45RB/RO binding chimeric antibody-treated and control cultures. 
     CD45RB/RO Binding Chimeric Antibody but not Control mAb Treated T Cells Display a T Regulatory Cell (Treg) Phenotype 
     Increased expression of CD25 and the negative regulatory protein CTLA-4 (CD152) is a marker of Treg cells. Functional suppression of primary and secondary T cell responses by CD45RB/RO binding chimeric antibody may be due to the induction of Treg cells. To address this issue, T cells were activated by anti-CD3+CD28 mAbs and cultured in the presence of CD45RB/RO binding chimeric antibody or anti-LPS control mAb. The time course of CTLA-4 and CD25 expression reveals marked differences between controls and CD45RB/RO binding chimeric antibody-treated T cells on days 1 and 3 after secondary stimulation, indicating a Treg phenotype. 
     Intracellular CTLA-4 Expression is Sustained in the Presence of CD45RB/RO Binding Chimeric Antibody 
     It has been reported that substantial amounts of CTLA-4 can also be found intracellularly. Therefore, in parallel to surface CTLA-4 staining, intracellular CTLA-4 expression was analyzed. Moderate differences between T cell cultures were seen on day 4 after stimulation. After prolonged culture, however, high levels of intracellular CTLA-4 were sustained only in CD45RB/RO binding chimeric antibody-treated but not in control T cells. 
     CD45RB/RO Binding Chimeric Antibody-Treated T Cells Become Double Positive for CD4 and CD8 
     Following stimulation, T cells induce and upregulate the expression of several surface receptors, such as CD25, CD152 (CTLA-4), CD154 (CD40-Ligand) and others. In contrast, the level of expression of CD4 or CD8 is thought to stay relatively constant. We reproducibly observed a strong increase of both CD4 and CD8 antigens on CD45RB/RO binding chimeric antibody-treated but not on control Ab-treated T cells after activation. The emergence of a CD4/CD8 double-positive T cell population seems to be due to the upregulation of CD4 on the CD8+ subset and conversely, CD8 on the CD4+ subset. This contrasts with a moderately low percentage of double positive T cells in control cultures. 
     High IL-2 Receptor Alpha-Chain, but Very Low Beta-Chain Expression by CD45RB/RO Binding Chimeric Antibody-Treated T Cells 
     Treg cells are known to be constitutively positive for CD25, the IL-2 receptor alpha-chain. The regulation of other subunits of the trimeric IL-2 receptor on Treg cells is not known. Recently we have compared the expression of the beta-chain of IL-2 receptor, e.g. CD122, on T cells activated and propagated in the presence or absence of CD45RB/RO binding chimeric antibody. The results show that CD45RB/RO binding chimeric antibody-treated T cells have about ten-fold lower CD122 expression as compared to T cells in control cultures. This difference may indicate that Treg cells require factors other than IL-2 to proliferate. 
     Example 8 
     Sequences of the Invention (CDR Sequences of the Invention are Underlined) 
     SEQ ID NO:1 
     Part of the Amino Acid Sequence of Chimeric Light Chain 
       
     
       
         
           
               
            
               
                 DILLTQSPAILSVSPGERVSFSC RASQNIGTSIQ WYQQRTNGSPRLLIR S   
               
               
                   
               
               
                   SSESIS GIPSRFSGSGSGTDFTLSINSVESEDIADYYC QQSNTWPFT FGS 
               
               
                   
               
               
                 GTKLEIK 
               
            
           
         
       
     
     SEQ ID NO:2 
     Part of the Amino Acid Sequence of Chimeric Heavy Chain 
       
     
       
         
           
               
            
               
                 EVQLQQSGPELVKPGASVKMSCKASGYTFT NYIIH WVKQEPGQGLEWIG Y   
               
               
                   
               
               
                   FNPYNHGTKYNEKFKG RATLTADKSSNTAYMDLSSLTSEDSAIYYCAR SG   
               
               
                   
               
               
                   PYAWFDT WGQGTTVTVSS 
               
            
           
         
       
     
     SEQ ID NO:3 
     Amino Acid Sequence of Chimeric Light Chain 
       
     
       
         
           
               
            
               
                 DILLTQSPAILSVSPGERVSFSC RASQNIGTSIQ WYQQRTNGSPRLLIR S   
               
               
                   
               
               
                   SSESIS GIPSRFSGSGSGTDFTLSINSVESEDIADYYC QQSNTWPFT FGS 
               
               
                   
               
               
                 GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKV 
               
               
                   
               
               
                 DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG 
               
               
                   
               
               
                 LSSPVTKSFNRGEC 
               
            
           
         
       
     
     SEQ ID NO:4 
     Amino Acid Sequence of Chimeric Heavy Chain 
       
     
       
         
           
               
            
               
                 EVQLQQSGPELVKPGASVKMSCKASGYTFT NYIIH WVKQEPGQGLEWIG Y   
               
               
                   
               
               
                   FNPYNHGTKYNEKFKG RATLTADKSSNTAYMDLSSLTSEDSAIYYCAR SG   
               
               
                   
               
               
                   PYAWFDT WGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY 
               
               
                   
               
               
                 FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI 
               
               
                   
               
               
                 CNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD 
               
               
                   
               
               
                 TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST 
               
               
                   
               
               
                 YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY 
               
               
                   
               
               
                 TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD 
               
               
                   
               
               
                 SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 
               
            
           
         
       
     
     SEQ ID NO:5 
     Nucleotide Sequence Encoding a Polypeptide of SEQ ID NO:1 
       
     
       
         
           
               
            
               
                 GACATTCTGCTGACCCAGTCTCCAGCCATCCTGTCTGTGAGTCCAGGAGA 
               
               
                   
               
               
                 AAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAACATTGGCACAAGCATAC 
               
               
                   
               
               
                 AGTGGTATCAACAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAGGTCT 
               
               
                   
               
               
                 TCTTCTGAGTCTATCTCTGGGATCCCTTCCAGGTTTAGTGGCAGTGGATC 
               
               
                   
               
               
                 AGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCTGAAGATATTG 
               
               
                   
               
               
                 CAGATTATTACTGTCAACAAAGTAATACCTGGCCATTCACGTTCGGCTCG 
               
               
                   
               
               
                 GGGACCAAGCTTGAAATCAAA 
               
            
           
         
       
     
     SEQ ID NO:6 
     Nucleotide Sequence Encoding a Polypeptide of SEQ ID NO:2 
       
     
       
         
           
               
            
               
                 GAGGTGCAGCTGCAGCAGTCAGGACCTGAACTGGTAAAGCCTGGGGCTTC 
               
               
                   
               
               
                 AGTGAAGATGTCCTGCAAGGCCTCTGGATACACATTCACTAATTATATTA 
               
               
                   
               
               
                 TCCACTGGGTGAAGCAGGAGCCTGGTCAGGGCCTTGAATGGATTGGATAT 
               
               
                   
               
               
                 TTTAATCCTTACAATCATGGTACTAAGTACAATGAGAAGTTCAAAGGCAG 
               
               
                   
               
               
                 GGCCACACTAACTGCAGACAAATCCTCCAACACAGCCTACATGGACCTCA 
               
               
                   
               
               
                 GCAGCCTGACCTCTGAGGACTCTGCGATCTACTACTGTGCAAGATCAGGA 
               
               
                   
               
               
                 CCCTATGCCTGGTTTGACACCTGGGGCCAAGGGACCACGGTCACCGTCTC 
               
               
                   
               
               
                 CTCA 
               
            
           
         
       
     
     SEQ ID NO:7 
     Part of Amino Acid Sequence of Humanised Light Chain Designated humV2 (humV2=VLm) 
     
       
         
           
               
               
            
               
                   
                 DILLTQSPAT LSLSPGERAT FSC RASQNIG TSIQW YQQKT 
               
               
                   
                   
               
               
                   
                 NGAPRLLIR S SSESIS GIPS RFSGSGSGTD FTLTISSLEP 
               
               
                   
                   
               
               
                   
                 EDFAVYYC QQ SNTWPFT FGQ GTKLEIK 
               
            
           
         
       
     
     SEQ ID NO:8 
     Part of Amino Acid Sequence of Humanised Light Chain Designated humV1 (humV1=VLh) 
     
       
         
           
               
               
            
               
                   
                 DILLTQSPAT LSLSPGERAT LSC RASQNIG TSIQW YQQKP 
               
               
                   
                   
               
               
                   
                 GQAPRLLIR S SSESIS GIPS RFSGSGSGTD FTLTISSLEP 
               
               
                   
                   
               
               
                   
                 EDFAVYYC QQ SNTWPFT FGQ GTKLEIK 
               
            
           
         
       
     
     SEQ ID NO:9 
     Part of Amino Acid Sequence of Humanised Heavy Chain Designated VHE 
       
     
       
         
           
               
               
            
               
                   
                 EVQLVESGAE VKKPGASVKV SCKASGYTFT  NYIIH WVKQE 
               
               
                   
                   
               
               
                   
                 PGQGLEWIG Y FNPYNHGTKY NEKFKG RATL TANKSISTAY 
               
               
                   
                   
               
               
                   
                 MELSSLRSED TAVYYCAR SG PYAWFDT WGQ GTTVTVSS 
               
            
           
         
       
     
     SEQ ID NO:10 
     Part of Amino Acid Sequence of Humanised Heavy Chain Designated VHQ 
       
     
       
         
           
               
               
            
               
                   
                 QVQLVESGAE VKKPGASVKV SCKASGYTFT  NYIIH WVKQE 
               
               
                   
                   
               
               
                   
                 PGQGLEWIG Y FNPYNHGTKY NEKFKG RATL TANKSISTAY 
               
               
                   
                   
               
               
                   
                 MELSSLRSED TAVYYCAR SG PYAWFDT WGQ GTTVTVSS 
               
            
           
         
       
     
     SEQ ID NO:11 
     Nucleotide Sequence Encoding Amino Acid Sequence SEQ ID NO:9 
       
     
       
         
           
               
            
               
                 GAGGTGCAGCTGGTGGAGTCAGGAGCCGAAGTGAAAAAGCCTGGGGCTTC 
               
               
                   
               
               
                 AGTGAAGGTGTCCTGCAAGGCCTCTGGATACACATTCACTAATTATATTA 
               
               
                   
               
               
                 TCCACTGGGTGAAGCAGGAGCCTGGTCAGGGCCTTGAATGGATTGGATAT 
               
               
                   
               
               
                 TTTAATCCTTACAATCATGGTACTAAGTACAATGAGAAGTTCAAAGGCAG 
               
               
                   
               
               
                 GGCCACACTAACTGCAAACAAATCCATCAGCACAGCCTACATGGAGCTCA 
               
               
                   
               
               
                 GCAGCCTGCGCTCTGAGGACACTGCGGTCTACTACTGTGCAAGATCAGGA 
               
               
                   
               
               
                 CCCTATGCCTGGTTTGACACCTGGGGCCAAGGGACCACGGTCACCGTCTC 
               
               
                   
               
               
                 CTCA 
               
            
           
         
       
     
     SEQ ID NO:12 
     Nucleotide Sequence Encoding Amino Acid Sequence SEQ ID NO:10 
       
     
       
         
           
               
            
               
                 CAGGTGCAGCTGGTGGAGTCAGGAGCCGAAGTGAAAAAGCCTGGGGCTTC 
               
               
                   
               
               
                 AGTGAAGGTGTCCTGCAAGGCCTCTGGATACACATTCACTAATTATATTA 
               
               
                   
               
               
                 TCCACTGGGTGAAGCAGGAGCCTGGTCAGGGCCTTGAATGGATTGGATAT 
               
               
                   
               
               
                 TTTAATCCTTACAATCATGGTACTAAGTACAATGAGAAGTTCAAAGGCAG 
               
               
                   
               
               
                 GGCCACACTAACTGCAAACAAATCCATCAGCACAGCCTACATGGAGCTCA 
               
               
                   
               
               
                 GCAGCCTGCGCTCTGAGGACACTGCGGTCTACTACTGTGCAAGATCAGGA 
               
               
                   
               
               
                 CCCTATGCCTGGTTTGACACCTGGGGCCAAGGGACCACGGTCACCGTCTC 
               
               
                   
               
               
                 CTCA 
               
            
           
         
       
     
     SEQ ID NO:13 
     Nucleotide Sequence Encoding Amino Acid Sequence SEQ ID NO:7 
       
     
       
         
           
               
            
               
                 GACATTCTGCTGACCCAGTCTCCAGCCACCCTGTCTCTGAGTCCAGGAGA 
               
               
                   
               
               
                 AAGAGCCACTTTCTCCTGCAGGGCCAGTCAGAACATTGGCACAAGCATAC 
               
               
                   
               
               
                 AGTGGTATCAACAAAAAACAAATGGTGCTCCAAGGCTTCTCATAAGGTCT 
               
               
                   
               
               
                 TCTTCTGAGTCTATCTCTGGGATCCCTTCCAGGTTTAGTGGCAGTGGATC 
               
               
                   
               
               
                 AGGGACAGATTTTACTCTTACCATCAGCAGTCTGGAGCCTGAAGATTTTG 
               
               
                   
               
               
                 CAGTGTATTACTGTCAACAAAGTAATACCTGGCCATTCACGTTCGGCCAG 
               
               
                   
               
               
                 GGGACCAAGCTGGAGATCAAA 
               
            
           
         
       
     
     SEQ ID NO:14 
     Nucleotide Sequence Encoding Amino Acid Sequence SEQ ID NO:8 
       
     
       
         
           
               
            
               
                 GACATTCTGCTGACCCAGTCTCCAGCCACCCTGTCTCTGAGTCCAGGAGA 
               
               
                   
               
               
                 AAGAGCCACTCTCTCCTGCAGGGCCAGTCAGAACATTGGCACAAGCATAC 
               
               
                   
               
               
                 AGTGGTATCAACAAAAACCAGGTCAGGCTCCAAGGCTTCTCATAAGGTCT 
               
               
                   
               
               
                 TCTTCTGAGTCTATCTCTGGGATCCCTTCCAGGTTTAGTGGCAGTGGATC 
               
               
                   
               
               
                 AGGGACAGATTTTACTCTTACCATCAGCAGTCTGGAGCCTGAAGATTTTG 
               
               
                   
               
               
                 CAGTGTATTACTGTCAACAAAGTAATACCTGGCCATTCACGTTCGGCCAG 
               
               
                   
               
               
                 GGGACCAAGCTGGAGATCAAA 
               
            
           
         
       
     
     SEQ ID NO:15 
     Nucleotide Sequence of the Expression Vector HCMV-G1 HuAb-VHQ 
     (Complete DNA Sequence of a Humanised Heavy Chain Expression Vector Comprising SEQ ID NO:12 (VHQ) from 3921-4274) 
     
       
         
           
               
               
               
            
               
                    1 
                 AGCTTTTTGC AAAAGCCTAG GCCTCCAAAA AAGCCTCCTC ACTACTTCTG 
                   
               
               
                   
               
               
                   51 
                 GAATAGCTCA GAGGCCGAGG CGGCCTCGGC CTCTGCATAA ATAAAAAAAA 
               
               
                   
               
               
                  101 
                 TTAGTCAGCC ATGGGGCGGA GAATGGGCGG AACTGGGCGG AGTTAGGGGC 
               
               
                   
               
               
                  151 
                 GGGATGGGCG GAGTTAGGGG CGGGACTATG GTTGCTGACT AATTGAGATG 
               
               
                   
               
               
                  201 
                 CATGCTTTGC ATACTTCTGC CTGCTGGGGA GCCTGGTTGC TGACTAATTG 
               
               
                   
               
               
                  251 
                 AGATGCATGC TTTGCATACT TCTGCCTGCT GGGGAGCCTG GGGACTTTCC 
               
               
                   
               
               
                  301 
                 ACACCCTAAC TGACACACAT TCCACAGCTG CCTCGCGCGT TTCGGTGATG 
               
               
                   
               
               
                  351 
                 ACGGTGAAAA CCTCTGACAC ATGCAGCTCC CGGAGACGGT CACAGCTTGT 
               
               
                   
               
               
                  401 
                 CTGTAAGCGG ATGCCGGGAG CAGACAAGCC CGTCAGGGCG CGTCAGCGGG 
               
               
                   
               
               
                  451 
                 TGTTGGCGGG TGTCGGGGCG CAGCCATGAC CCAGTCACGT AGCGATAGCG 
               
               
                   
               
               
                  501 
                 GAGTGTATAC TGGCTTAACT ATGCGGCATC AGAGCAGATT GTACTGAGAG 
               
               
                   
               
               
                  551 
                 TGCACCATAT GCGGTGTGAA ATACCGCACA GATGCGTAAG GAGAAAATAC 
               
               
                   
               
               
                  601 
                 CGCATCAGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT 
               
               
                   
               
               
                  651 
                 CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT 
               
               
                   
               
               
                  701 
                 TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC 
               
               
                   
               
               
                  751 
                 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA 
               
               
                   
               
               
                  801 
                 TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA 
               
               
                   
               
               
                  851 
                 GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA 
               
               
                   
               
               
                  901 
                 AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT 
               
               
                   
               
               
                  951 
                 GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT AGCTCACGCT 
               
               
                   
               
               
                 1001 
                 GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG 
               
               
                   
               
               
                 1051 
                 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG 
               
               
                   
               
               
                 1101 
                 TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA 
               
               
                   
               
               
                 1151 
                 CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC 
               
               
                   
               
               
                 1201 
                 TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGGACAG TATTTGGTAT 
               
               
                   
               
               
                 1251 
                 CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT GGTAGCTCTT 
               
               
                   
               
               
                 1301 
                 GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG 
               
               
                   
               
               
                 1351 
                 CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT 
               
               
                   
               
               
                 1401 
                 TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT 
               
               
                   
               
               
                 1451 
                 TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA 
               
               
                   
               
               
                 1501 
                 AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA 
               
               
                   
               
               
                 1551 
                 CAGTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT 
               
               
                   
               
               
                 1601 
                 TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT AACTACGATA 
               
               
                   
               
               
                 1651 
                 CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC 
               
               
                   
               
               
                 1701 
                 ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG 
               
               
                   
               
               
                 1751 
                 CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT 
               
               
                   
               
               
                 1801 
                 AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG 
               
               
                   
               
               
                 1851 
                 CAACGTTGTT GCCATTGCTG CAGGCATCGT GGTGTCACGC TCGTCGTTTG 
               
               
                   
               
               
                 1901 
                 GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG AGTTACATGA 
               
               
                   
               
               
                 1951 
                 TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT 
               
               
                   
               
               
                 2001 
                 TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC 
               
               
                   
               
               
                 2051 
                 TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT 
               
               
                   
               
               
                 2101 
                 GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG 
               
               
                   
               
               
                 2151 
                 TTGCTCTTGC CCGGCGTCAA CACGGGATAA TACCGCGCCA CATAGCAGAA 
               
               
                   
               
               
                 2201 
                 CTTTAAAAGT GCTCATCATT GGAAAACGTT CTTCGGGGCG AAAACTCTCA 
               
               
                   
               
               
                 2251 
                 AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC 
               
               
                   
               
               
                 2301 
                 CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA 
               
               
                   
               
               
                 2351 
                 AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA 
               
               
                   
               
               
                 2401 
                 TGTTGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA 
               
               
                   
               
               
                 2451 
                 GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA 
               
               
                   
               
               
                 2501 
                 AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC ACCTGACGTC 
               
               
                   
               
               
                 2551 
                 TAAGAAACCA TTATTATCAT GACATTAACC TATAAAAATA GGCGTATCAC 
               
               
                   
               
               
                 2601 
                 GAGGCCCTTT CGTCTTCAAG AATTCAGCTT GGCTGCAGTG AATAATAAAA 
               
               
                   
               
               
                 2651 
                 TGTGTGTTTG TCCGAAATAC GCGTTTTGAG ATTTCTGTCG CCGACTAAAT 
               
               
                   
               
               
                 2701 
                 TCATGTCGCG CGATAGTGGT GTTTATCGCC GATAGAGATG GCGATATTGG 
               
               
                   
               
               
                 2751 
                 AAAAATCGAT ATTTGAAAAT ATGGCATATT GAAAATGTCG CCGATGTGAG 
               
               
                   
               
               
                 2801 
                 TTTCTGTGTA ACTGATATCG CCATTTTTCC AAAAGTGATT TTTGGGCATA 
               
               
                   
               
               
                 2851 
                 CGCGATATCT GGCGATAGCG CTTATATCGT TTACGGGGGA TGGCGATAGA 
               
               
                   
               
               
                 2901 
                 CGACTTTGGT GACTTGGGCG ATTCTGTGTG TCGCAAATAT CGCAGTTTCG 
               
               
                   
               
               
                 2951 
                 ATATAGGTGA CAGACGATAT GAGGCTATAT CGCCGATAGA GGCGACATCA 
               
               
                   
               
               
                 3001 
                 AGCTGGCACA TGGCCAATGC ATATCGATCT ATACATTGAA TCAATATTGG 
               
               
                   
               
               
                 3051 
                 CCATTAGCCA TATTATTCAT TGGTTATATA GCATAAATCA ATATTGGCTA 
               
               
                   
               
               
                 3101 
                 TTGGCCATTG CATACGTTGT ATCCATATCA TAATATGTAC ATTTATATTG 
               
               
                   
               
               
                 3151 
                 GCTCATGTCC AACATTACCG CCATGTTGAC ATTGATTATT GACTAGTTAT 
               
               
                   
               
               
                 3201 
                 TAATAGTAAT CAATTACGGG GTCATTAGTT CATAGCCCAT ATATGGAGTT 
               
               
                   
               
               
                 3251 
                 CCGCGTTACA TAACTTACGG TAAATGGCCC GCCTGGCTGA CCGCCCAACG 
               
               
                   
               
               
                 3301 
                 ACCCCCGCCC ATTGACGTCA ATAATGACGT ATGTTCCCAT AGTAACGCCA 
               
               
                   
               
               
                 3351 
                 ATAGGGACTT TCCATTGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC 
               
               
                   
               
               
                 3401 
                 CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTACG CCCCCTATTG 
               
               
                   
               
               
                 3451 
                 ACGTCAATGA CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC 
               
               
                   
               
               
                 3501 
                 TTATGGGACT TTCCTACTTG GCAGTACATC TACGTATTAG TCATCGCTAT 
               
               
                   
               
               
                 3551 
                 TACCATGGTG ATGCGGTTTT GGCAGTACAT CAATGGGCGT GGATAGCGGT 
               
               
                   
               
               
                 3601 
                 TTGACTCACG GGGATTTCCA AGTCTCCACC CCATTGACGT CAATGGGAGT 
               
               
                   
               
               
                 3651 
                 TTGTTTTGGC ACCAAAATCA ACGGGACTTT CCAAAATGTC GTAACAACTC 
               
               
                   
               
               
                 3701 
                 CGCCCCATTG ACGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA 
               
               
                   
               
               
                 3751 
                 TAAGCAGAGC TCGTTTAGTG AACCGTCAGA TCGCCTGGAG ACGCCATCCA 
               
               
                   
               
               
                 3801 
                 CGCTGTTTTG ACCTCCATAG AAGACACCGG GACCGATCCA GCCTCCGCAA 
               
               
                   
               
               
                 3851 
                 GCTTGCCGCC ACCATGGACT GGACCTGGAG GGTGTTCTGC CTGCTGGCCG 
               
               
                   
               
               
                 3901 
                 TGGCCCCCGG CGCCCACAGC CAGGTGCAGC TGGTGGAGTC AGGAGCCGAA 
               
               
                   
               
               
                 3951 
                 GTGAAAAAGC CTGGGGCTTC AGTGAAGGTG TCCTGCAAGG CCTCTGGATA 
               
               
                   
               
               
                 4001 
                 CACATTCACT AATTATATTA TCCACTGGGT GAAGCAGGAG CCTGGTCAGG 
               
               
                   
               
               
                 4051 
                 GCCTTGAATG GATTGGATAT TTTAATCCTT ACAATCATGG TACTAAGTAC 
               
               
                   
               
               
                 4101 
                 AATGAGAAGT TCAAAGGCAG GGCCACACTA ACTGCAAACA AATCCATCAG 
               
               
                   
               
               
                 4151 
                 CACAGCCTAC ATGGAGCTCA GCAGCCTGCG CTCTGAGGAC ACTGCGGTCT 
               
               
                   
               
               
                 4201 
                 ACTACTGTGC AAGATCAGGA CCCTATGCCT GGTTTGACAC CTGGGGCCAA 
               
               
                   
               
               
                 4251 
                 GGGACCACGG TCACCGTCTC CTCAGGTGAG TTCTAGAAGG ATCCCAAGCT 
               
               
                   
               
               
                 4301 
                 AGCTTTCTGG GGCAGGCCAG GCCTGACCTT GGCTTTGGGG CAGGGAGGGG 
               
               
                   
               
               
                 4351 
                 GCTAAGGTGA GGCAGGTGGC GCCAGCCAGG TGCACACCCA ATGCCCATGA 
               
               
                   
               
               
                 4401 
                 GCCCAGACAC TGGACGCTGA ACCTCGCGGA CAGTTAAGAA CCCAGGGGCC 
               
               
                   
               
               
                 4451 
                 TCTGCGCCCT GGGCCCAGCT CTGTCCCACA CCGCGGTCAC ATGGCACCAC 
               
               
                   
               
               
                 4501 
                 CTCTCTTGCA GCCTCCACCA AGGGCCCATC GGTCTTCCCC CTGGCACCCT 
               
               
                   
               
               
                 4551 
                 CCTCCAAGAG CACCTCTGGG GGCACAGCGG CCCTGGGCTG CCTGGTCAAG 
               
               
                   
               
               
                 4601 
                 GACTACTTCC CCGAACCGGT GACGGTGTCG TGGAACTCAG GCGCCCTGAC 
               
               
                   
               
               
                 4651 
                 CAGCGGCGTG CACACCTTCC CGGCTGTCCT ACAGTCCTCA GGACTCTACT 
               
               
                   
               
               
                 4701 
                 CCCTCAGCAG CGTGGTGACC GTGCCCTCCA GCAGCTTGGG CACCCAGACC 
               
               
                   
               
               
                 4751 
                 TACATCTGCA ACGTGAATCA CAAGCCCAGC AACACCAAGG TGGACAAGAA 
               
               
                   
               
               
                 4801 
                 AGTTGGTGAG AGGCCAGCAC AGGGAGGGAG GGTGTCTGCT GGAAGCCAGG 
               
               
                   
               
               
                 4851 
                 CTCAGCGCTC CTGCCTGGAC GCATCCCGGC TATGCAGCCC CAGTCCAGGG 
               
               
                   
               
               
                 4901 
                 CAGCAAGGCA GGCCCCGTCT GCCTCTTCAC CCGGAGGCCT CTGCCCGCCC 
               
               
                   
               
               
                 4951 
                 CACTCATGCT CAGGGAGAGG GTCTTCTGGC TTTTTCCCCA GGCTCTGGGC 
               
               
                   
               
               
                 5001 
                 AGGCACAGGC TAGGTGCCCC TAACCCAGGC CCTGCACACA AAGGGGCAGG 
               
               
                   
               
               
                 5051 
                 TGCTGGGCTC AGACCTGCCA AGAGCCATAT CCGGGAGGAC CCTGCCCCTG 
               
               
                   
               
               
                 5101 
                 ACCTAAGCCC ACCCCAAAGG CCAAACTCTC CACTCCCTCA GCTCGGACAC 
               
               
                   
               
               
                 5151 
                 CTTCTCTCCT CCCAGATTCC AGTAACTCCC AATCTTCTCT CTGCAGAGCC 
               
               
                   
               
               
                 5201 
                 CAAATCTTGT GACAAAACTC ACACATGCCC ACCGTGCCCA GGTAAGCCAG 
               
               
                   
               
               
                 5251 
                 CCCAGGCCTC GCCCTCCAGC TCAAGGCGGG ACAGGTGCCC TAGAGTAGCC 
               
               
                   
               
               
                 5301 
                 TGCATCCAGG GACAGGCCCC AGCCGGGTGC TGACACGTCC ACCTCCATCT 
               
               
                   
               
               
                 5351 
                 CTTCCTCAGC ACCTGAACTC CTGGGGGGAC CGTCAGTCTT CCTCTTCCCC 
               
               
                   
               
               
                 5401 
                 CCAAAACCCA AGGACACCCT CATGATCTCC CGGACCCCTG AGGTCACATG 
               
               
                   
               
               
                 5451 
                 CGTGGTGGTG GACGTGAGCC ACGAAGACCC TGAGGTCAAG TTCAACTGGT 
               
               
                   
               
               
                 5501 
                 ACGTGGACGG CGTGGAGGTG CATAATGCCA AGACAAAGCC GCGGGAGGAG 
               
               
                   
               
               
                 5551 
                 CAGTACAACA GCACGTACCG TGTGGTCAGC GTCCTCACCG TCCTGCACCA 
               
               
                   
               
               
                 5601 
                 GGACTGGCTG AATGGCAAGG AGTACAAGTG CAAGGTCTCC AACAAAGCCC 
               
               
                   
               
               
                 5651 
                 TCCCAGCCCC CATCGAGAAA ACCATCTCCA AAGCCAAAGG TGGGACCCGT 
               
               
                   
               
               
                 5701 
                 GGGGTGCGAG GGCCACATGG ACAGAGGCCG GCTCGGCCCA CCCTCTGCCC 
               
               
                   
               
               
                 5751 
                 TGAGAGTGAC CGCTGTACCA ACCTCTGTCC CTACAGGGCA GCCCCGAGAA 
               
               
                   
               
               
                 5801 
                 CCACAGGTGT ACACCCTGCC CCCATCCCGG GATGAGCTGA CCAAGAACCA 
               
               
                   
               
               
                 5851 
                 GGTCAGCCTG ACCTGCCTGG TCAAAGGCTT CTATCCCAGC GACATCGCCG 
               
               
                   
               
               
                 5901 
                 TGGAGTGGGA GAGCAATGGG CAGCCGGAGA ACAACTACAA GACCACGCCT 
               
               
                   
               
               
                 5951 
                 CCCGTGCTGG ACTCCGACGG CTCCTTCTTC CTCTACAGCA AGCTCACCGT 
               
               
                   
               
               
                 6001 
                 GGACAAGAGC AGGTGGCAGC AGGGGAACGT CTTCTCATGC TCCGTGATGC 
               
               
                   
               
               
                 6051 
                 ATGAGGCTCT GCACAACCAC TACACGCAGA AGAGCCTCTC CCTGTCTCCG 
               
               
                   
               
               
                 6101 
                 GGTAAATGAG TGCGACGGCC GGCAAGCCCC CGCTCCCCGG GCTCTCGCGG 
               
               
                   
               
               
                 6151 
                 TCGCACGAGG ATGCTTGGCA CGTACCCCCT GTACATACTT CCCGGGCGCC 
               
               
                   
               
               
                 6201 
                 CAGCATGGAA ATAAAGCACC CAGCGCTGCC CTGGGCCCCT GCGAGACTGT 
               
               
                   
               
               
                 6251 
                 GATGGTTCTT TCCACGGGTC AGGCCGAGTC TGAGGCCTGA GTGGCATGAG 
               
               
                   
               
               
                 6301 
                 ATCTGATATC ATCGATGAAT TCGAGCTCGG TACCCGGGGA TCGATCCAGA 
               
               
                   
               
               
                 6351 
                 CATGATAAGA TACATTGATG AGTTTGGACA AACCACAACT AGAATGCAGT 
               
               
                   
               
               
                 6401 
                 GAAAAAAATG CTTTATTTGT GAAATTTGTG ATGCTATTGC TTTATTTGTA 
               
               
                   
               
               
                 6451 
                 ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT GCATTCATTT 
               
               
                   
               
               
                 6501 
                 TATGTTTCAG GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA AGCAAGTAAA 
               
               
                   
               
               
                 6551 
                 ACCTCTACAA ATGTGGTATG GCTGATTATG ATCTCTAGTC AAGGCACTAT 
               
               
                   
               
               
                 6601 
                 ACATCAAATA TTCCTTATTA ACCCCTTTAC AAATTAAAAA GCTAAAGGTA 
               
               
                   
               
               
                 6651 
                 CACAATTTTT GAGCATAGTT ATTAATAGCA GACACTCTAT GCCTGTGTGG 
               
               
                   
               
               
                 6701 
                 AGTAAGAAAA AACAGTATGT TATGATTATA ACTGTTATGC CTACTTATAA 
               
               
                   
               
               
                 6751 
                 AGGTTACAGA ATATTTTTCC ATAATTTTCT TGTATAGCAG TGCAGCTTTT 
               
               
                   
               
               
                 6801 
                 TCCTTTGTGG TGTAAATAGC AAAGCAAGCA AGAGTTCTAT TACTAAACAC 
               
               
                   
               
               
                 6851 
                 AGCATGACTC AAAAAACTTA GCAATTCTGA AGGAAAGTCC TTGGGGTCTT 
               
               
                   
               
               
                 6901 
                 CTACCTTTCT CTTCTTTTTT GGAGGAGTAG AATGTTGAGA GTCAGCAGTA 
               
               
                   
               
               
                 6951 
                 GCCTCATCAT CACTAGATGG CATTTCTTCT GAGCAAAACA GGTTTTCCTC 
               
               
                   
               
               
                 7001 
                 ATTAAAGGCA TTCCACCACT GCTCCCATTC ATCAGTTCCA TAGGTTGGAA 
               
               
                   
               
               
                 7051 
                 TCTAAAATAC ACAAACAATT AGAATCAGTA GTTTAACACA TTATACACTT 
               
               
                   
               
               
                 7101 
                 AAAAATTTTA TATTTACCTT AGAGCTTTAA ATCTCTGTAG GTAGTTTGTC 
               
               
                   
               
               
                 7151 
                 CAATTATGTC ACACCACAGA AGTAAGGTTC CTTCACAAAG ATCCGGGACC 
               
               
                   
               
               
                 7201 
                 AAAGCGGCCA TCGTGCCTCC CCACTCCTGC AGTTCGGGGG CATGGATGCG 
               
               
                   
               
               
                 7251 
                 CGGATAGCCG CTGCTGGTTT CCTGGATGCC GACGGATTTG CACTGCCGGT 
               
               
                   
               
               
                 7301 
                 AGAACTCCGC GAGGTCGTCC AGCCTCAGGC AGCAGCTGAA CCAACTCGCG 
               
               
                   
               
               
                 7351 
                 AGGGGATCGA GCCCGGGGTG GGCGAAGAAC TCCAGCATGA GATCCCCGCG 
               
               
                   
               
               
                 7401 
                 CTGGAGGATC ATCCAGCCGG CGTCCCGGAA AACGATTCCG AAGCCCAACC 
               
               
                   
               
               
                 7451 
                 TTTCATAGAA GGCGGCGGTG GAATCGAAAT CTCGTGATGG CAGGTTGGGC 
               
               
                   
               
               
                 7501 
                 GTCGCTTGGT CGGTCATTTC GAACCCCAGA GTCCCGCTCA GAAGAACTCG 
               
               
                   
               
               
                 7551 
                 TCAAGAAGGC GATAGAAGGC GATGCGCTGC GAATCGGGAG CGGCGATACC 
               
               
                   
               
               
                 7601 
                 GTAAAGCACG AGGAAGCGGT CAGCCCATTC GCCGCCAAGC TCTTCAGCAA 
               
               
                   
               
               
                 7651 
                 TATCACGGGT AGCCAACGCT ATGTCCTGAT AGCGGTCCGC CACACCCAGC 
               
               
                   
               
               
                 7701 
                 CGGCCACAGT CGATGAATCC AGAAAAGCGG CCATTTTCCA CCATGATATT 
               
               
                   
               
               
                 7751 
                 CGGCAAGCAG GCATCGCCAT GGGTCACGAC GAGATCCTCG CCGTCGGGCA 
               
               
                   
               
               
                 7801 
                 TGCGCGCCTT GAGCCTGGCG AACAGTTCGG CTGGCGCGAG CCCCTGATGC 
               
               
                   
               
               
                 7851 
                 TCTTCGTCCA GATCATCCTG ATCGACAAGA CCGGCTTCCA TCCGAGTACG 
               
               
                   
               
               
                 7901 
                 TGCTCGCTCG ATGCGATGTT TCGCTTGGTG GTCGAATGGG CAGGTAGCCG 
               
               
                   
               
               
                 7951 
                 GATCAAGCGT ATGCAGCCGC CGCATTGCAT CAGCCATGAT GGATACTTTC 
               
               
                   
               
               
                 8001 
                 TCGGCAGGAG CAAGGTGAGA TGACAGGAGA TCCTGCCCCG GCACTTCGCC 
               
               
                   
               
               
                 8051 
                 CAATAGCAGC CAGTCCCTTC CCGCTTCAGT GACAACGTCG AGCACAGCTG 
               
               
                   
               
               
                 8101 
                 CGCAAGGAAC GCCCGTCGTG GCCAGCCACG ATAGCCGCGC TGCCTCGTCC 
               
               
                   
               
               
                 8151 
                 TGCAGTTCAT TCAGGGCACC GGACAGGTCG GTCTTGACAA AAAGAACCGG 
               
               
                   
               
               
                 8201 
                 GCGCCCCTGC GCTGACAGCC GGAACACGGC GGCATCAGAG CAGCCGATTG 
               
               
                   
               
               
                 8251 
                 TCTGTTGTGC CCAGTCATAG CCGAATAGCC TCTCCACCCA AGCGGCCGGA 
               
               
                   
               
               
                 8301 
                 GAACCTGCGT GCAATCCATC TTGTTCAATC ATGCGAAACG ATCCTCATCC 
               
               
                   
               
               
                 8351 
                 TGTCTCTTGA TCAGATCTTG ATCCCCTGCG CCATCAGATC CTTGGCGGCA 
               
               
                   
               
               
                 8401 
                 AGAAAGCCAT CCAGTTTACT TTGCAGGGCT TCCCAACCTT ACCAGAGGGC 
               
               
                   
               
               
                 8451 
                 GCCCCAGCTG GCAATTCCGG TTCGCTTGCT GTCCATAAAA CCGCCCAGTC 
               
               
                   
               
               
                 8501 
                 TAGCTATCGC CATGTAAGCC CACTGCAAGC TACCTGCTTT CTCTTTGCGC 
               
               
                   
               
               
                 8551 
                 TTGCGTTTTC CCTTGTCCAG ATAGCCCAGT AGCTGACATT CATCCGGGGT 
               
               
                   
               
               
                 8601 
                 CAGCACCGTT TCTGCGGACT GGCTTTCTAC GTGTTCCGCT TCCTTTAGCA 
               
               
                   
               
               
                 8651 
                 GCCCTTGCGC CCTGAGTGCT TGCGGCAGCG TGAAGCT 
               
            
           
         
       
     
     SEQ ID NO:16 
     Nucleotide Sequence of the Expression Vector HCMV-G1 HuAb-VHE 
     (Complete DNA Sequence of a Humanised Heavy Chain Expression Vector Comprising SEQ ID NO: 11 (VHE) from 3921-4274) 
     
       
         
           
               
               
               
            
               
                    1 
                 AGCTTTTTGC AAAAGCCTAG GCCTCCAAAA AAGCCTCCTC ACTACTTCTG 
                   
               
               
                   
               
               
                   51 
                 GAATAGCTCA GAGGCCGAGG CGGCCTCGGC CTCTGCATAA ATAAAAAAAA 
               
               
                   
               
               
                  101 
                 TTAGTCAGCC ATGGGGCGGA GAATGGGCGG AACTGGGCGG AGTTAGGGGC 
               
               
                   
               
               
                  151 
                 GGGATGGGCG GAGTTAGGGG CGGGACTATG GTTGCTGACT AATTGAGATG 
               
               
                   
               
               
                  201 
                 CATGCTTTGC ATACTTCTGC CTGCTGGGGA GCCTGGTTGC TGACTAATTG 
               
               
                   
               
               
                  251 
                 AGATGCATGC TTTGCATACT TCTGCCTGCT GGGGAGCCTG GGGACTTTCC 
               
               
                   
               
               
                  301 
                 ACACCCTAAC TGACACACAT TCCACAGCTG CCTCGCGCGT TTCGGTGATG 
               
               
                   
               
               
                  351 
                 ACGGTGAAAA CCTCTGACAC ATGCAGCTCC CGGAGACGGT CACAGCTTGT 
               
               
                   
               
               
                  401 
                 CTGTAAGCGG ATGCCGGGAG CAGACAAGCC CGTCAGGGCG CGTCAGCGGG 
               
               
                   
               
               
                  451 
                 TGTTGGCGGG TGTCGGGGCG CAGCCATGAC CCAGTCACGT AGCGATAGCG 
               
               
                   
               
               
                  501 
                 GAGTGTATAC TGGCTTAACT ATGCGGCATC AGAGCAGATT GTACTGAGAG 
               
               
                   
               
               
                  551 
                 TGCACCATAT GCGGTGTGAA ATACCGCACA GATGCGTAAG GAGAAAATAC 
               
               
                   
               
               
                  601 
                 CGCATCAGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT 
               
               
                   
               
               
                  651 
                 CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT 
               
               
                   
               
               
                  701 
                 TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC 
               
               
                   
               
               
                  751 
                 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA 
               
               
                   
               
               
                  801 
                 TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA 
               
               
                   
               
               
                  851 
                 GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA 
               
               
                   
               
               
                  901 
                 AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT 
               
               
                   
               
               
                  951 
                 GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT AGCTCACGCT 
               
               
                   
               
               
                 1001 
                 GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG 
               
               
                   
               
               
                 1051 
                 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG 
               
               
                   
               
               
                 1101 
                 TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA 
               
               
                   
               
               
                 1151 
                 CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC 
               
               
                   
               
               
                 1201 
                 TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGGACAG TATTTGGTAT 
               
               
                   
               
               
                 1251 
                 CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT GGTAGCTCTT 
               
               
                   
               
               
                 1301 
                 GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG 
               
               
                   
               
               
                 1351 
                 CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT 
               
               
                   
               
               
                 1401 
                 TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT 
               
               
                   
               
               
                 1451 
                 TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA 
               
               
                   
               
               
                 1501 
                 AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA 
               
               
                   
               
               
                 1551 
                 CAGTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT 
               
               
                   
               
               
                 1601 
                 TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT AACTACGATA 
               
               
                   
               
               
                 1651 
                 CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC 
               
               
                   
               
               
                 1701 
                 ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG 
               
               
                   
               
               
                 1751 
                 CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT 
               
               
                   
               
               
                 1801 
                 AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG 
               
               
                   
               
               
                 1851 
                 CAACGTTGTT GCCATTGCTG CAGGCATCGT GGTGTCACGC TCGTCGTTTG 
               
               
                   
               
               
                 1901 
                 GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG AGTTACATGA 
               
               
                   
               
               
                 1951 
                 TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT 
               
               
                   
               
               
                 2001 
                 TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC 
               
               
                   
               
               
                 2051 
                 TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT 
               
               
                   
               
               
                 2101 
                 GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG 
               
               
                   
               
               
                 2151 
                 TTGCTCTTGC CCGGCGTCAA CACGGGATAA TACCGCGCCA CATAGCAGAA 
               
               
                   
               
               
                 2201 
                 CTTTAAAAGT GCTCATCATT GGAAAACGTT CTTCGGGGCG AAAACTCTCA 
               
               
                   
               
               
                 2251 
                 AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC 
               
               
                   
               
               
                 2301 
                 CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA 
               
               
                   
               
               
                 2351 
                 AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA 
               
               
                   
               
               
                 2401 
                 TGTTGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA 
               
               
                   
               
               
                 2451 
                 GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA 
               
               
                   
               
               
                 2501 
                 AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC ACCTGACGTC 
               
               
                   
               
               
                 2551 
                 TAAGAAACCA TTATTATCAT GACATTAACC TATAAAAATA GGCGTATCAC 
               
               
                   
               
               
                 2601 
                 GAGGCCCTTT CGTCTTCAAG AATTCAGCTT GGCTGCAGTG AATAATAAAA 
               
               
                   
               
               
                 2651 
                 TGTGTGTTTG TCCGAAATAC GCGTTTTGAG ATTTCTGTCG CCGACTAAAT 
               
               
                   
               
               
                 2701 
                 TCATGTCGCG CGATAGTGGT GTTTATCGCC GATAGAGATG GCGATATTGG 
               
               
                   
               
               
                 2751 
                 AAAAATCGAT ATTTGAAAAT ATGGCATATT GAAAATGTCG CCGATGTGAG 
               
               
                   
               
               
                 2801 
                 TTTCTGTGTA ACTGATATCG CCATTTTTCC AAAAGTGATT TTTGGGCATA 
               
               
                   
               
               
                 2851 
                 CGCGATATCT GGCGATAGCG CTTATATCGT TTACGGGGGA TGGCGATAGA 
               
               
                   
               
               
                 2901 
                 CGACTTTGGT GACTTGGGCG ATTCTGTGTG TCGCAAATAT CGCAGTTTCG 
               
               
                   
               
               
                 2951 
                 ATATAGGTGA CAGACGATAT GAGGCTATAT CGCCGATAGA GGCGACATCA 
               
               
                   
               
               
                 3001 
                 AGCTGGCACA TGGCCAATGC ATATCGATCT ATACATTGAA TCAATATTGG 
               
               
                   
               
               
                 3051 
                 CCATTAGCCA TATTATTCAT TGGTTATATA GCATAAATCA ATATTGGCTA 
               
               
                   
               
               
                 3101 
                 TTGGCCATTG CATACGTTGT ATCCATATCA TAATATGTAC ATTTATATTG 
               
               
                   
               
               
                 3151 
                 GCTCATGTCC AACATTACCG CCATGTTGAC ATTGATTATT GACTAGTTAT 
               
               
                   
               
               
                 3201 
                 TAATAGTAAT CAATTACGGG GTCATTAGTT CATAGCCCAT ATATGGAGTT 
               
               
                   
               
               
                 3251 
                 CCGCGTTACA TAACTTACGG TAAATGGCCC GCCTGGCTGA CCGCCCAACG 
               
               
                   
               
               
                 3301 
                 ACCCCCGCCC ATTGACGTCA ATAATGACGT ATGTTCCCAT AGTAACGCCA 
               
               
                   
               
               
                 3351 
                 ATAGGGACTT TCCATTGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC 
               
               
                   
               
               
                 3401 
                 CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTACG CCCCCTATTG 
               
               
                   
               
               
                 3451 
                 ACGTCAATGA CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC 
               
               
                   
               
               
                 3501 
                 TTATGGGACT TTCCTACTTG GCAGTACATC TACGTATTAG TCATCGCTAT 
               
               
                   
               
               
                 3551 
                 TACCATGGTG ATGCGGTTTT GGCAGTACAT CAATGGGCGT GGATAGCGGT 
               
               
                   
               
               
                 3601 
                 TTGACTCACG GGGATTTCCA AGTCTCCACC CCATTGACGT CAATGGGAGT 
               
               
                   
               
               
                 3651 
                 TTGTTTTGGC ACCAAAATCA ACGGGACTTT CCAAAATGTC GTAACAACTC 
               
               
                   
               
               
                 3701 
                 CGCCCCATTG ACGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA 
               
               
                   
               
               
                 3751 
                 TAAGCAGAGC TCGTTTAGTG AACCGTCAGA TCGCCTGGAG ACGCCATCCA 
               
               
                   
               
               
                 3801 
                 CGCTGTTTTG ACCTCCATAG AAGACACCGG GACCGATCCA GCCTCCGCAA 
               
               
                   
               
               
                 3851 
                 GCTTGCCGCC ACCATGGACT GGACCTGGAG GGTGTTCTGC CTGCTGGCCG 
               
               
                   
               
               
                 3901 
                 TGGCCCCCGG CGCCCACAGC GAGGTGCAGC TGGTGGAGTC AGGAGCCGAA 
               
               
                   
               
               
                 3951 
                 GTGAAAAAGC CTGGGGCTTC AGTGAAGGTG TCCTGCAAGG CCTCTGGATA 
               
               
                   
               
               
                 4001 
                 CACATTCACT AATTATATTA TCCACTGGGT GAAGCAGGAG CCTGGTCAGG 
               
               
                   
               
               
                 4051 
                 GCCTTGAATG GATTGGATAT TTTAATCCTT ACAATCATGG TACTAAGTAC 
               
               
                   
               
               
                 4101 
                 AATGAGAAGT TCAAAGGCAG GGCCACACTA ACTGCAAACA AATCCATCAG 
               
               
                   
               
               
                 4151 
                 CACAGCCTAC ATGGAGCTCA GCAGCCTGCG CTCTGAGGAC ACTGCGGTCT 
               
               
                   
               
               
                 4201 
                 ACTACTGTGC AAGATCAGGA CCCTATGCCT GGTTTGACAC CTGGGGCCAA 
               
               
                   
               
               
                 4251 
                 GGGACCACGG TCACCGTCTC CTCAGGTGAG TTCTAGAAGG ATCCCAAGCT 
               
               
                   
               
               
                 4301 
                 AGCTTTCTGG GGCAGGCCAG GCCTGACCTT GGCTTTGGGG CAGGGAGGGG 
               
               
                   
               
               
                 4351 
                 GCTAAGGTGA GGCAGGTGGC GCCAGCCAGG TGCACACCCA ATGCCCATGA 
               
               
                   
               
               
                 4401 
                 GCCCAGACAC TGGACGCTGA ACCTCGCGGA CAGTTAAGAA CCCAGGGGCC 
               
               
                   
               
               
                 4451 
                 TCTGCGCCCT GGGCCCAGCT CTGTCCCACA CCGCGGTCAC ATGGCACCAC 
               
               
                   
               
               
                 4501 
                 CTCTCTTGCA GCCTCCACCA AGGGCCCATC GGTCTTCCCC CTGGCACCCT 
               
               
                   
               
               
                 4551 
                 CCTCCAAGAG CACCTCTGGG GGCACAGCGG CCCTGGGCTG CCTGGTCAAG 
               
               
                   
               
               
                 4601 
                 GACTACTTCC CCGAACCGGT GACGGTGTCG TGGAACTCAG GCGCCCTGAC 
               
               
                   
               
               
                 4651 
                 CAGCGGCGTG CACACCTTCC CGGCTGTCCT ACAGTCCTCA GGACTCTACT 
               
               
                   
               
               
                 4701 
                 CCCTCAGCAG CGTGGTGACC GTGCCCTCCA GCAGCTTGGG CACCCAGACC 
               
               
                   
               
               
                 4751 
                 TACATCTGCA ACGTGAATCA CAAGCCCAGC AACACCAAGG TGGACAAGAA 
               
               
                   
               
               
                 4801 
                 AGTTGGTGAG AGGCCAGCAC AGGGAGGGAG GGTGTCTGCT GGAAGCCAGG 
               
               
                   
               
               
                 4851 
                 CTCAGCGCTC CTGCCTGGAC GCATCCCGGC TATGCAGCCC CAGTCCAGGG 
               
               
                   
               
               
                 4901 
                 CAGCAAGGCA GGCCCCGTCT GCCTCTTCAC CCGGAGGCCT CTGCCCGCCC 
               
               
                   
               
               
                 4951 
                 CACTCATGCT CAGGGAGAGG GTCTTCTGGC TTTTTCCCCA GGCTCTGGGC 
               
               
                   
               
               
                 5001 
                 AGGCACAGGC TAGGTGCCCC TAACCCAGGC CCTGCACACA AAGGGGCAGG 
               
               
                   
               
               
                 5051 
                 TGCTGGGCTC AGACCTGCCA AGAGCCATAT CCGGGAGGAC CCTGCCCCTG 
               
               
                   
               
               
                 5101 
                 ACCTAAGCCC ACCCCAAAGG CCAAACTCTC CACTCCCTCA GCTCGGACAC 
               
               
                   
               
               
                 5151 
                 CTTCTCTCCT CCCAGATTCC AGTAACTCCC AATCTTCTCT CTGCAGAGCC 
               
               
                   
               
               
                 5201 
                 CAAATCTTGT GACAAAACTC ACACATGCCC ACCGTGCCCA GGTAAGCCAG 
               
               
                   
               
               
                 5251 
                 CCCAGGCCTC GCCCTCCAGC TCAAGGCGGG ACAGGTGCCC TAGAGTAGCC 
               
               
                   
               
               
                 5301 
                 TGCATCCAGG GACAGGCCCC AGCCGGGTGC TGACACGTCC ACCTCCATCT 
               
               
                   
               
               
                 5351 
                 CTTCCTCAGC ACCTGAACTC CTGGGGGGAC CGTCAGTCTT CCTCTTCCCC 
               
               
                   
               
               
                 5401 
                 CCAAAACCCA AGGACACCCT CATGATCTCC.CGGACCCCTG AGGTCACATG 
               
               
                   
               
               
                 5451 
                 CGTGGTGGTG GACGTGAGCC ACGAAGACCC TGAGGTCAAG TTCAACTGGT 
               
               
                   
               
               
                 5501 
                 ACGTGGACGG CGTGGAGGTG CATAATGCCA AGACAAAGCC GCGGGAGGAG 
               
               
                   
               
               
                 5551 
                 CAGTACAACA GCACGTACCG TGTGGTCAGC GTCCTCACCG TCCTGCACCA 
               
               
                   
               
               
                 5601 
                 GGACTGGCTG AATGGCAAGG AGTACAAGTG CAAGGTCTCC AACAAAGCCC 
               
               
                   
               
               
                 5651 
                 TCCCAGCCCC CATCGAGAAA ACCATCTCCA AAGCCAAAGG TGGGACCCGT 
               
               
                   
               
               
                 5701 
                 GGGGTGCGAG GGCCACATGG ACAGAGGCCG GCTCGGCCCA CCCTCTGCCC 
               
               
                   
               
               
                 5751 
                 TGAGAGTGAC CGCTGTACCA ACCTCTGTCC CTACAGGGCA GCCCCGAGAA 
               
               
                   
               
               
                 5801 
                 CCACAGGTGT ACACCCTGCC CCCATCCCGG GATGAGCTGA CCAAGAACCA 
               
               
                   
               
               
                 5851 
                 GGTCAGCCTG ACCTGCCTGG TCAAAGGCTT CTATCCCAGC GACATCGCCG 
               
               
                   
               
               
                 5901 
                 TGGAGTGGGA GAGCAATGGG CAGCCGGAGA ACAACTACAA GACCACGCCT 
               
               
                   
               
               
                 5951 
                 CCCGTGCTGG ACTCCGACGG CTCCTTCTTC CTCTACAGCA AGCTCACCGT 
               
               
                   
               
               
                 6001 
                 GGACAAGAGC AGGTGGCAGC AGGGGAACGT CTTCTCATGC TCCGTGATGC 
               
               
                   
               
               
                 6051 
                 ATGAGGCTCT GCACAACCAC TACACGCAGA AGAGCCTCTC CCTGTCTCCG 
               
               
                   
               
               
                 6101 
                 GGTAAATGAG TGCGACGGCC GGCAAGCCCC CGCTCCCCGG GCTCTCGCGG 
               
               
                   
               
               
                 6151 
                 TCGCACGAGG ATGCTTGGCA CGTACCCCCT GTACATACTT CCCGGGCGCC 
               
               
                   
               
               
                 6201 
                 CAGCATGGAA ATAAAGCACC CAGCGCTGCC CTGGGCCCCT GCGAGACTGT 
               
               
                   
               
               
                 6251 
                 GATGGTTCTT TCCACGGGTC AGGCCGAGTC TGAGGCCTGA GTGGCATGAG 
               
               
                   
               
               
                 6301 
                 ATCTGATATC ATCGATGAAT TCGAGCTCGG TACCCGGGGA TCGATCCAGA 
               
               
                   
               
               
                 6351 
                 CATGATAAGA TACATTGATG AGTTTGGACA AACCACAACT AGAATGCAGT 
               
               
                   
               
               
                 6401 
                 GAAAAAAATG CTTTATTTGT GAAATTTGTG ATGCTATTGC TTTATTTGTA 
               
               
                   
               
               
                 6451 
                 ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT GCATTCATTT 
               
               
                   
               
               
                 6501 
                 TATGTTTCAG GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA AGCAAGTAAA 
               
               
                   
               
               
                 6551 
                 ACCTCTACAA ATGTGGTATG GCTGATTATG ATCTCTAGTC AAGGCACTAT 
               
               
                   
               
               
                 6601 
                 ACATCAAATA TTCCTTATTA ACCCCTTTAC AAATTAAAAA GCTAAAGGTA 
               
               
                   
               
               
                 6651 
                 CACAATTTTT GAGCATAGTT ATTAATAGCA GACACTCTAT GCCTGTGTGG 
               
               
                   
               
               
                 6701 
                 AGTAAGAAAA AACAGTATGT TATGATTATA ACTGTTATGC CTACTTATAA 
               
               
                   
               
               
                 6751 
                 AGGTTACAGA ATATTTTTCC ATAATTTTCT TGTATAGCAG TGCAGCTTTT 
               
               
                   
               
               
                 6801 
                 TCCTTTGTGG TGTAAATAGC AAAGCAAGCA AGAGTTCTAT TACTAAACAC 
               
               
                   
               
               
                 6851 
                 AGCATGACTC AAAAAACTTA GCAATTCTGA AGGAAAGTCC TTGGGGTCTT 
               
               
                   
               
               
                 6901 
                 CTACCTTTCT CTTCTTTTTT GGAGGAGTAG AATGTTGAGA GTCAGCAGTA 
               
               
                   
               
               
                 6951 
                 GCCTCATCAT CACTAGATGG CATTTCTTCT GAGCAAAACA GGTTTTCCTC 
               
               
                   
               
               
                 7001 
                 ATTAAAGGCA TTCCACCACT GCTCCCATTC ATCAGTTCCA TAGGTTGGAA 
               
               
                   
               
               
                 7051 
                 TCTAAAATAC ACAAACAATT AGAATCAGTA GTTTAACACA TTATACACTT 
               
               
                   
               
               
                 7101 
                 AAAAATTTTA TATTTACCTT AGAGCTTTAA ATCTCTGTAG GTAGTTTGTC 
               
               
                   
               
               
                 7151 
                 CAATTATGTC ACACCACAGA AGTAAGGTTC CTTCACAAAG ATCCGGGACC 
               
               
                   
               
               
                 7201 
                 AAAGCGGCCA TCGTGCCTCC CCACTCCTGC AGTTCGGGGG CATGGATGCG 
               
               
                   
               
               
                 7251 
                 CGGATAGCCG CTGCTGGTTT CCTGGATGCC GACGGATTTG CACTGCCGGT 
               
               
                   
               
               
                 7301 
                 AGAACTCCGC GAGGTCGTCC AGCCTCAGGC AGCAGCTGAA CCAACTCGCG 
               
               
                   
               
               
                 7351 
                 AGGGGATCGA GCCCGGGGTG GGCGAAGAAC TCCAGCATGA GATCCCCGCG 
               
               
                   
               
               
                 7401 
                 CTGGAGGATC ATCCAGCCGG CGTCCCGGAA AACGATTCCG AAGCCCAACC 
               
               
                   
               
               
                 7451 
                 TTTCATAGAA GGCGGCGGTG GAATCGAAAT CTCGTGATGG CAGGTTGGGC 
               
               
                   
               
               
                 7501 
                 GTCGCTTGGT CGGTCATTTC GAACCCCAGA GTCCCGCTCA GAAGAACTCG 
               
               
                   
               
               
                 7551 
                 TCAAGAAGGC GATAGAAGGC GATGCGCTGC GAATCGGGAG CGGCGATACC 
               
               
                   
               
               
                 7601 
                 GTAAAGCACG AGGAAGCGGT CAGCCCATTC GCCGCCAAGC TCTTCAGCAA 
               
               
                   
               
               
                 7651 
                 TATCACGGGT AGCCAACGCT ATGTCCTGAT AGCGGTCCGC CACACCCAGC 
               
               
                   
               
               
                 7701 
                 CGGCCACAGT CGATGAATCC AGAAAAGCGG CCATTTTCCA CCATGATATT 
               
               
                   
               
               
                 7751 
                 CGGCAAGCAG GCATCGCCAT GGGTCACGAC GAGATCCTCG CCGTCGGGCA 
               
               
                   
               
               
                 7801 
                 TGCGCGCCTT GAGCCTGGCG AACAGTTCGG CTGGCGCGAG CCCCTGATGC 
               
               
                   
               
               
                 7851 
                 TCTTCGTCCA GATCATCCTG ATCGACAAGA CCGGCTTCCA TCCGAGTACG 
               
               
                   
               
               
                 7901 
                 TGCTCGCTCG ATGCGATGTT TCGCTTGGTG GTCGAATGGG CAGGTAGCCG 
               
               
                   
               
               
                 7951 
                 GATCAAGCGT ATGCAGCCGC CGCATTGCAT CAGCCATGAT GGATACTTTC 
               
               
                   
               
               
                 8001 
                 TCGGCAGGAG CAAGGTGAGA TGACAGGAGA TCCTGCCCCG GCACTTCGCC 
               
               
                   
               
               
                 8051 
                 CAATAGCAGC CAGTCCCTTC CCGCTTCAGT GACAACGTCG AGCACAGCTG 
               
               
                   
               
               
                 8101 
                 CGCAAGGAAC GCCCGTCGTG GCCAGCCACG ATAGCCGCGC TGCCTCGTCC 
               
               
                   
               
               
                 8151 
                 TGCAGTTCAT TCAGGGCACC GGACAGGTCG GTCTTGACAA AAAGAACCGG 
               
               
                   
               
               
                 8201 
                 GCGCCCCTGC GCTGACAGCC GGAACACGGC GGCATCAGAG CAGCCGATTG 
               
               
                   
               
               
                 8251 
                 TCTGTTGTGC CCAGTCATAG CCGAATAGCC TCTCCACCCA AGCGGCCGGA 
               
               
                   
               
               
                 8301 
                 GAACCTGCGT GCAATCCATC TTGTTCAATC ATGCGAAACG ATCCTCATCC 
               
               
                   
               
               
                 8351 
                 TGTCTCTTGA TCAGATCTTG ATCCCCTGCG CCATCAGATC CTTGGCGGCA 
               
               
                   
               
               
                 8401 
                 AGAAAGCCAT CCAGTTTACT TTGCAGGGCT TCCCAACCTT ACCAGAGGGC 
               
               
                   
               
               
                 8451 
                 GCCCCAGCTG GCAATTCCGG TTCGCTTGCT GTCCATAAAA CCGCCCAGTC 
               
               
                   
               
               
                 8501 
                 TAGCTATCGC CATGTAAGCC CACTGCAAGC TACCTGCTTT CTCTTTGCGC 
               
               
                   
               
               
                 8551 
                 TTGCGTTTTC CCTTGTCCAG ATAGCCCAGT AGCTGACATT CATCCGGGGT 
               
               
                   
               
               
                 8601 
                 CAGCACCGTT TCTGCGGACT GGCTTTCTAC GTGTTCCGCT TCCTTTAGCA 
               
               
                   
               
               
                 8651 
                 GCCCTTGCGC CCTGAGTGCT TGCGGCAGCG TGAAGCT 
               
            
           
         
       
     
     SEQ ID NO:17 
     Nucleotide Sequence of the Expression Vector HCMV-K HuAb-VL1 hum V1 
     (Complete DNA Sequence of a Humanised Light Chain Expression Vector Comprising SEQ ID NO: 14 (humV1=VLh) from 3964-4284) 
     
       
         
           
               
               
               
            
               
                    1 
                 CTAGCTTTTT GCAAAAGCCT AGGCCTCCAA AAAAGCCTCC TCACTACTTC 
                   
               
               
                   
               
               
                   51 
                 TGGAATAGCT CAGAGGCCGA GGCGGCCTCG GCCTCTGCAT AAATAAAAAA 
               
               
                   
               
               
                  101 
                 AATTAGTCAG CCATGGGGCG GAGAATGGGC GGAACTGGGC GGAGTTAGGG 
               
               
                   
               
               
                  151 
                 GCGGGATGGG CGGAGTTAGG GGCGGGACTA TGGTTGCTGA CTAATTGAGA 
               
               
                   
               
               
                  201 
                 TGCATGCTTT GCATACTTCT GCCTGCTGGG GAGCCTGGTT GCTGACTAAT 
               
               
                   
               
               
                  251 
                 TGAGATGCAT GCTTTGCATA CTTCTGCCTG CTGGGGAGCC TGGGGACTTT 
               
               
                   
               
               
                  301 
                 CCACACCCTA ACTGACACAC ATTCCACAGC TGCCTCGCGC GTTTCGGTGA 
               
               
                   
               
               
                  351 
                 TGACGGTGAA AACCTCTGAC ACATGCAGCT CCCGGAGACG GTCACAGCTT 
               
               
                   
               
               
                  401 
                 GTCTGTAAGC GGATGCCGGG AGCAGACAAG CCCGTCAGGG CGCGTCAGCG 
               
               
                   
               
               
                  451 
                 GGTGTTGGCG GGTGTCGGGG CGCAGCCATG ACCCAGTCAC GTAGCGATAG 
               
               
                   
               
               
                  501 
                 CGGAGTGTAT ACTGGCTTAA CTATGCGGCA TCAGAGCAGA TTGTACTGAG 
               
               
                   
               
               
                  551 
                 AGTGCACCAT ATGCGGTGTG AAATACCGCA CAGATGCGTA AGGAGAAAAT 
               
               
                   
               
               
                  601 
                 ACCGCATCAG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG 
               
               
                   
               
               
                  651 
                 GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG 
               
               
                   
               
               
                  701 
                 GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG 
               
               
                   
               
               
                  751 
                 GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC 
               
               
                   
               
               
                  801 
                 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA 
               
               
                   
               
               
                  851 
                 GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG 
               
               
                   
               
               
                  901 
                 GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC 
               
               
                   
               
               
                  951 
                 CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG 
               
               
                   
               
               
                 1001 
                 CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG 
               
               
                   
               
               
                 1051 
                 TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT 
               
               
                   
               
               
                 1101 
                 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC 
               
               
                   
               
               
                 1151 
                 CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT 
               
               
                   
               
               
                 1201 
                 TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGGAC AGTATTTGGT 
               
               
                   
               
               
                 1251 
                 ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC 
               
               
                   
               
               
                 1301 
                 TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA 
               
               
                   
               
               
                 1351 
                 AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC 
               
               
                   
               
               
                 1401 
                 TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT 
               
               
                   
               
               
                 1451 
                 TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT 
               
               
                   
               
               
                 1501 
                 AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT 
               
               
                   
               
               
                 1551 
                 GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT 
               
               
                   
               
               
                 1601 
                 ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA 
               
               
                   
               
               
                 1651 
                 TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC 
               
               
                   
               
               
                 1701 
                 CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG 
               
               
                   
               
               
                 1751 
                 GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA 
               
               
                   
               
               
                 1801 
                 TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG 
               
               
                   
               
               
                 1851 
                 CGCAACGTTG TTGCCATTGC TGCAGGCATC GTGGTGTCAC GCTCGTCGTT 
               
               
                   
               
               
                 1901 
                 TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT 
               
               
                   
               
               
                 1951 
                 GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC 
               
               
                   
               
               
                 2001 
                 GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC 
               
               
                   
               
               
                 2051 
                 ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA 
               
               
                   
               
               
                 2101 
                 CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG 
               
               
                   
               
               
                 2151 
                 AGTTGCTCTT GCCCGGCGTC AACACGGGAT AATACCGCGC CACATAGCAG 
               
               
                   
               
               
                 2201 
                 AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT 
               
               
                   
               
               
                 2251 
                 CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA 
               
               
                   
               
               
                 2301 
                 CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC 
               
               
                   
               
               
                 2351 
                 AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA 
               
               
                   
               
               
                 2401 
                 AATGTTGAAT ACTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT 
               
               
                   
               
               
                 2451 
                 CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA 
               
               
                   
               
               
                 2501 
                 TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG 
               
               
                   
               
               
                 2551 
                 TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC 
               
               
                   
               
               
                 2601 
                 ACGAGGCCCT TTCGTCTTCA AGAATTCAGC TTGGCTGCAG TGAATAATAA 
               
               
                   
               
               
                 2651 
                 AATGTGTGTT TGTCCGAAAT ACGCGTTTTG AGATTTCTGT CGCCGACTAA 
               
               
                   
               
               
                 2701 
                 ATTCATGTCG CGCGATAGTG GTGTTTATCG CCGATAGAGA TGGCGATATT 
               
               
                   
               
               
                 2751 
                 GGAAAAATCG ATATTTGAAA ATATGGCATA TTGAAAATGT CGCCGATGTG 
               
               
                   
               
               
                 2801 
                 AGTTTCTGTG TAACTGATAT CGCCATTTTT CCAAAAGTGA TTTTTGGGCA 
               
               
                   
               
               
                 2851 
                 TACGCGATAT CTGGCGATAG CGCTTATATC GTTTACGGGG GATGGCGATA 
               
               
                   
               
               
                 2901 
                 GACGACTTTG GTGACTTGGG CGATTCTGTG TGTCGCAAAT ATCGCAGTTT 
               
               
                   
               
               
                 2951 
                 CGATATAGGT GACAGACGAT ATGAGGCTAT ATCGCCGATA GAGGCGACAT 
               
               
                   
               
               
                 3001 
                 CAAGCTGGCA CATGGCCAAT GCATATCGAT CTATACATTG AATCAATATT 
               
               
                   
               
               
                 3051 
                 GGCCATTAGC CATATTATTC ATTGGTTATA TAGCATAAAT CAATATTGGC 
               
               
                   
               
               
                 3101 
                 TATTGGCCAT TGCATACGTT GTATCCATAT CATAATATGT ACATTTATAT 
               
               
                   
               
               
                 3151 
                 TGGCTCATGT CCAACATTAC CGCCATGTTG ACATTGATTA TTGACTAGTT 
               
               
                   
               
               
                 3201 
                 ATTAATAGTA ATCAATTACG GGGTCATTAG TTCATAGCCC ATATATGGAG 
               
               
                   
               
               
                 3251 
                 TTCCGCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT GACCGCCCAA 
               
               
                   
               
               
                 3301 
                 CGACCCCCGC CCATTGACGT CAATAATGAC GTATGTTCCC ATAGTAACGC 
               
               
                   
               
               
                 3351 
                 CAATAGGGAC TTTCCATTGA CGTCAATGGG TGGAGTATTT ACGGTAAACT 
               
               
                   
               
               
                 3401 
                 GCCCACTTGG CAGTACATCA AGTGTATCAT ATGCCAAGTA CGCCCCCTAT 
               
               
                   
               
               
                 3451 
                 TGACGTCAAT GACGGTAAAT GGCCCGCCTG GCATTATGCC CAGTACATGA 
               
               
                   
               
               
                 3501 
                 CCTTATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT AGTCATCGCT 
               
               
                   
               
               
                 3551 
                 ATTACCATGG TGATGCGGTT TTGGCAGTAC ATCAATGGGC GTGGATAGCG 
               
               
                   
               
               
                 3601 
                 GTTTGACTCA CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA 
               
               
                   
               
               
                 3651 
                 GTTTGTTTTG GCACCAAAAT CAACGGGACT TTCCAAAATG TCGTAACAAC 
               
               
                   
               
               
                 3701 
                 TCCGCCCCAT TGACGCAAAT GGGCGGTAGG CGTGTACGGT GGGAGGTCTA 
               
               
                   
               
               
                 3751 
                 TATAAGCAGA GCTCGTTTAG TGAACCGTCA GATCGCCTGG AGACGCCATC 
               
               
                   
               
               
                 3801 
                 CACGCTGTTT TGACCTCCAT AGAAGACACC GGGACCGATC CAGCCTCCGC 
               
               
                   
               
               
                 3851 
                 AAGCTTGATA TCGAATTCCT GCAGCCCGGG GGATCCGCCC GCTTGCCGCC 
               
               
                   
               
               
                 3901 
                 ACCATGGAGA CCCCCGCCCA GCTGCTGTTC CTGCTGCTGC TGTGGCTGCC 
               
               
                   
               
               
                 3951 
                 CGACACCACC GGCGACATTC TGCTGACCCA GTCTCCAGCC ACCCTGTCTC 
               
               
                   
               
               
                 4001 
                 TGAGTCCAGG AGAAAGAGCC ACTCTCTCCT GCAGGGCCAG TCAGAACATT 
               
               
                   
               
               
                 4051 
                 GGCACAAGCA TACAGTGGTA TCAACAAAAA CCAGGTCAGG CTCCAAGGCT 
               
               
                   
               
               
                 4101 
                 TCTCATAAGG TCTTCTTCTG AGTCTATCTC TGGGATCCCT TCCAGGTTTA 
               
               
                   
               
               
                 4151 
                 GTGGCAGTGG ATCAGGGACA GATTTTACTC TTACCATCAG CAGTCTGGAG 
               
               
                   
               
               
                 4201 
                 CCTGAAGATT TTGCAGTGTA TTACTGTCAA CAAAGTAATA CCTGGCCATT 
               
               
                   
               
               
                 4251 
                 CACGTTCGGC CAGGGGACCA AGCTGGAGAT CAAACGTGAG TATTCTAGAA 
               
               
                   
               
               
                 4301 
                 AGATCCTAGA ATTCTAAACT CTGAGGGGGT CGGATGACGT GGCCATTCTT 
               
               
                   
               
               
                 4351 
                 TGCCTAAAGC ATTGAGTTTA CTGCAAGGTC AGAAAAGCAT GCAAAGCCCT 
               
               
                   
               
               
                 4401 
                 CAGAATGGCT GCAAAGAGCT CCAACAAAAC AATTTAGAAC TTTATTAAGG 
               
               
                   
               
               
                 4451 
                 AATAGGGGGA AGCTAGGAAG AAACTCAAAA CATCAAGATT TTAAATACGC 
               
               
                   
               
               
                 4501 
                 TTCTTGGTCT CCTTGCTATA ATTATCTGGG ATAAGCATGC TGTTTTCTGT 
               
               
                   
               
               
                 4551 
                 CTGTCCCTAA CATGCCCTGT GATTATCCGC AAACAACACA CCCAAGGGCA 
               
               
                   
               
               
                 4601 
                 GAACTTTGTT ACTTAAACAC CATCCTGTTT GCTTCTTTCC TCAGGAACTG 
               
               
                   
               
               
                 4651 
                 TGGCTGCACC ATCTGTCTTC ATCTTCCCGC CATCTGATGA GCAGTTGAAA 
               
               
                   
               
               
                 4701 
                 TCTGGAACTG CCTCTGTTGT GTGCCTGCTG AATAACTTCT ATCCCAGAGA 
               
               
                   
               
               
                 4751 
                 GGCCAAAGTA CAGTGGAAGG TGGATAACGC CCTCCAATCG GGTAACTCCC 
               
               
                   
               
               
                 4801 
                 AGGAGAGTGT CACAGAGCAG GACAGCAAGG ACAGCACCTA CAGCCTCAGC 
               
               
                   
               
               
                 4851 
                 AGCACCCTGA CGCTGAGCAA AGCAGACTAC GAGAAACACA AAGTCTACGC 
               
               
                   
               
               
                 4901 
                 CTGCGAAGTC ACCCATCAGG GCCTGAGCTC GCCCGTCACA AAGAGCTTCA 
               
               
                   
               
               
                 4951 
                 ACAGGGGAGA GTGTTAGAGG GAGAAGTGCC CCCACCTGCT CCTCAGTTCC 
               
               
                   
               
               
                 5001 
                 AGCCTGACCC CCTCCCATCC TTTGGCCTCT GACCCTTTTT CCACAGGGGA 
               
               
                   
               
               
                 5051 
                 CCTACCCCTA TTGCGGTCCT CCAGCTCATC TTTCACCTCA CCCCCCTCCT 
               
               
                   
               
               
                 5101 
                 CCTCCTTGGC TTTAATTATG CTAATGTTGG AGGAGAATGA ATAAATAAAG 
               
               
                   
               
               
                 5151 
                 TGAATCTTTG CACCTGTGGT TTCTCTCTTT CCTCATTTAA TAATTATTAT 
               
               
                   
               
               
                 5201 
                 CTGTTGTTTA CCAACTACTC AATTTCTCTT ATAAGGGACT AAATATGTAG 
               
               
                   
               
               
                 5251 
                 TCATCCTAAG GCGCATAACC ATTTATAAAA ATCATCCTTC ATTCTATTTT 
               
               
                   
               
               
                 5301 
                 ACCCTATCAT CCTCTGCAAG ACAGTCCTCC CTCAAACCCA CAAGCCTTCT 
               
               
                   
               
               
                 5351 
                 GTCCTCACAG TCCCCTGGGC CATGGTAGGA GAGACTTGCT TCCTTGTTTT 
               
               
                   
               
               
                 5401 
                 CCCCTCCTCA GCAAGCCCTC ATAGTCCTTT TTAAGGGTGA CAGGTCTTAC 
               
               
                   
               
               
                 5451 
                 AGTCATATAT CCTTTGATTC AATTCCCTGA GAATCAACCA AAGCAAATTT 
               
               
                   
               
               
                 5501 
                 TTCAAAAGAA GAAACCTGCT ATAAAGAGAA TCATTCATTG CAACATGATA 
               
               
                   
               
               
                 5551 
                 TAAAATAACA ACACAATAAA AGCAATTAAA TAAACAAACA ATAGGGAAAT 
               
               
                   
               
               
                 5601 
                 GTTTAAGTTC ATCATGGTAC TTAGACTTAA TGGAATGTCA TGCCTTATTT 
               
               
                   
               
               
                 5651 
                 ACATTTTTAA ACAGGTACTG AGGGACTCCT GTCTGCCAAG GGCCGTATTG 
               
               
                   
               
               
                 5701 
                 AGTACTTTCC ACAACCTAAT TTAATCCACA CTATACTGTG AGATTAAAAA 
               
               
                   
               
               
                 5751 
                 CATTCATTAA AATGTTGCAA AGGTTCTATA AAGCTGAGAG ACAAATATAT 
               
               
                   
               
               
                 5801 
                 TCTATAACTC AGCAATCCCA CTTCTAGATG ACTGAGTGTC CCCACCCACC 
               
               
                   
               
               
                 5851 
                 AAAAAACTAT GCAAGAATGT TCAAAGCAGC TTTATTTACA AAAGCCAAAA 
               
               
                   
               
               
                 5901 
                 ATTGGAAATA GCCCGATTGT CCAACAATAG AATGAGTTAT TAAACTGTGG 
               
               
                   
               
               
                 5951 
                 TATGTTTATA CATTAGAATA CCCAATGAGG AGAATTAACA AGCTACAACT 
               
               
                   
               
               
                 6001 
                 ATACCTACTC ACACAGATGA ATCTCATAAA AATAATGTTA CATAAGAGAA 
               
               
                   
               
               
                 6051 
                 ACTCAATGCA AAAGATATGT TCTGTATGTT TTCATCCATA TAAAGTTCAA 
               
               
                   
               
               
                 6101 
                 AACCAGGTAA AAATAAAGTT AGAAATTTGG ATGGAAATTA CTCTTAGCTG 
               
               
                   
               
               
                 6151 
                 GGGGTGGGCG AGTTAGTGCC TGGGAGAAGA CAAGAAGGGG CTTCTGGGGT 
               
               
                   
               
               
                 6201 
                 CTTGGTAATG TTCTGTTCCT CGTGTGGGGT TGTGCAGTTA TGATCTGTGC 
               
               
                   
               
               
                 6251 
                 ACTGTTCTGT ATACACATTA TGCTTCAAAA TAACTTCACA TAAAGAACAT 
               
               
                   
               
               
                 6301 
                 CTTATACCCA GTTAATAGAT AGAAGAGGAA TAAGTAATAG GTCAAGACCA 
               
               
                   
               
               
                 6351 
                 CGCAGCTGGT AAGTGGGGGG GCCTGGGATC AAATAGCTAC CTGCCTAATC 
               
               
                   
               
               
                 6401 
                 CTGCCCTCTT GAGCCCTGAA TGAGTCTGCC TTCCAGGGCT CAAGGTGCTC 
               
               
                   
               
               
                 6451 
                 AACAAAACAA CAGGCCTGCT ATTTTCCTGG CATCTGTGCC CTGTTTGGCT 
               
               
                   
               
               
                 6501 
                 AGCTAGGAGC ACACATACAT AGAAATTAAA TGAAACAGAC CTTCAGCAAG 
               
               
                   
               
               
                 6551 
                 GGGACAGAGG ACAGAATTAA CCTTGCCCAG ACACTGGAAA CCCATGTATG 
               
               
                   
               
               
                 6601 
                 AACACTCACA TGTTTGGGAA GGGGGAAGGG CACATGTAAA TGAGGACTCT 
               
               
                   
               
               
                 6651 
                 TCCTCATTCT ATGGGGCACT CTGGCCCTGC CCCTCTCAGC TACTCATCCA 
               
               
                   
               
               
                 6701 
                 TCCAACACAC CTTTCTAAGT ACCTCTCTCT GCCTACACTC TGAAGGGGTT 
               
               
                   
               
               
                 6751 
                 CAGGAGTAAC TAACACAGCA TCCCTTCCCT CAAATGACTG ACAATCCCTT 
               
               
                   
               
               
                 6801 
                 TGTCCTGCTT TGTTTTTCTT TCCAGTCAGT ACTGGGAAAG TGGGGAAGGA 
               
               
                   
               
               
                 6851 
                 CAGTCATGGA GAAACTACAT AAGGAAGCAC CTTGCCCTTC TGCCTCTTGA 
               
               
                   
               
               
                 6901 
                 GAATGTTGAT GAGTATCAAA TCTTTCAAAC TTTGGAGGTT TGAGTAGGGG 
               
               
                   
               
               
                 6951 
                 TGAGACTCAG TAATGTCCCT TCCAATGACA TGAACTTGCT CACTCATCCC 
               
               
                   
               
               
                 7001 
                 TGGGGGCCAA ATTGAACAAT CAAAGGCAGG CATAATCCAG CTATGAATTC 
               
               
                   
               
               
                 7051 
                 TAGGATCGAT CCAGACATGA TAAGATACAT TGATGAGTTT GGACAAACCA 
               
               
                   
               
               
                 7101 
                 CAACTAGAAT GCAGTGAAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT 
               
               
                   
               
               
                 7151 
                 ATTGCTTTAT TTGTAACCAT TATAAGCTGC AATAAACAAG TTAACAACAA 
               
               
                   
               
               
                 7201 
                 CAATTGCATT CATTTTATGT TTCAGGTTCA GGGGGAGGTG TGGGAGGTTT 
               
               
                   
               
               
                 7251 
                 TTTAAAGCAA GTAAAACCTC TACAAATGTG GTATGGCTGA TTATGATCTC 
               
               
                   
               
               
                 7301 
                 TAGTCAAGGC ACTATACATC AAATATTCCT TATTAACCCC TTTACAAATT 
               
               
                   
               
               
                 7351 
                 AAAAAGCTAA AGGTACACAA TTTTTGAGCA TAGTTATTAA TAGCAGACAC 
               
               
                   
               
               
                 7401 
                 TCTATGCCTG TGTGGAGTAA GAAAAAACAG TATGTTATGA TTATAACTGT 
               
               
                   
               
               
                 7451 
                 TATGCCTACT TATAAAGGTT ACAGAATATT TTTCCATAAT TTTCTTGTAT 
               
               
                   
               
               
                 7501 
                 AGCAGTGCAG CTTTTTCCTT TGTGGTGTAA ATAGCAAAGC AAGCAAGAGT 
               
               
                   
               
               
                 7551 
                 TCTATTACTA AACACAGCAT GACTCAAAAA ACTTAGCAAT TCTGAAGGAA 
               
               
                   
               
               
                 7601 
                 AGTCCTTGGG GTCTTCTACC TTTCTCTTCT TTTTTGGAGG AGTAGAATGT 
               
               
                   
               
               
                 7651 
                 TGAGAGTCAG CAGTAGCCTC ATCATCACTA GATGGCATTT CTTCTGAGCA 
               
               
                   
               
               
                 7701 
                 AAACAGGTTT TCCTCATTAA AGGCATTCCA CCACTGCTCC CATTCATCAG 
               
               
                   
               
               
                 7751 
                 TTCCATAGGT TGGAATCTAA AATACACAAA CAATTAGAAT CAGTAGTTTA 
               
               
                   
               
               
                 7801 
                 ACACATTATA CACTTAAAAA TTTTATATTT ACCTTAGAGC TTTAAATCTC 
               
               
                   
               
               
                 7851 
                 TGTAGGTAGT TTGTCCAATT ATGTCACACC ACAGAAGTAA GGTTCCTTCA 
               
               
                   
               
               
                 7901 
                 CAAAGATCCG GGACCAAAGC GGCCATCGTG CCTCCCCACT CCTGCAGTTC 
               
               
                   
               
               
                 7951 
                 GGGGGCATGG ATGCGCGGAT AGCCGCTGCT GGTTTCCTGG ATGCCGACGG 
               
               
                   
               
               
                 8001 
                 ATTTGCACTG CCGGTAGAAC TCCGCGAGGT CGTCCAGCCT CAGGCAGCAG 
               
               
                   
               
               
                 8051 
                 CTGAACCAAC TCGCGAGGGG ATCGAGCCCG GGGTGGGCGA AGAACTCCAG 
               
               
                   
               
               
                 8101 
                 CATGAGATCC CCGCGCTGGA GGATCATCCA GCCGGCGTCC CGGAAAACGA 
               
               
                   
               
               
                 8151 
                 TTCCGAAGCC CAACCTTTCA TAGAAGGCGG CGGTGGAATC GAAATCTCGT 
               
               
                   
               
               
                 8201 
                 GATGGCAGGT TGGGCGTCGC TTGGTCGGTC ATTTCGAACC CCAGAGTCCC 
               
               
                   
               
               
                 8251 
                 GCTCAGAAGA ACTCGTCAAG AAGGCGATAG AAGGCGATGC GCTGCGAATC 
               
               
                   
               
               
                 8301 
                 GGGAGCGGCG ATACCGTAAA GCACGAGGAA GCGGTCAGCC CATTCGCCGC 
               
               
                   
               
               
                 8351 
                 CAAGCTCTTC AGCAATATCA CGGGTAGCCA ACGCTATGTC CTGATAGCGG 
               
               
                   
               
               
                 8401 
                 TCCGCCACAC CCAGCCGGCC ACAGTCGATG AATCCAGAAA AGCGGCCATT 
               
               
                   
               
               
                 8451 
                 TTCCACCATG ATATTCGGCA AGCAGGCATC GCCATGGGTC ACGACGAGAT 
               
               
                   
               
               
                 8501 
                 CCTCGCCGTC GGGCATGCGC GCCTTGAGCC TGGCGAACAG TTCGGCTGGC 
               
               
                   
               
               
                 8551 
                 GCGAGCCCCT GATGCTCTTC GTCCAGATCA TCCTGATCGA CAAGACCGGC 
               
               
                   
               
               
                 8601 
                 TTCCATCCGA GTACGTGCTC GCTCGATGCG ATGTTTCGCT TGGTGGTCGA 
               
               
                   
               
               
                 8651 
                 ATGGGCAGGT AGCCGGATCA AGCGTATGCA GCCGCCGCAT TGCATCAGCC 
               
               
                   
               
               
                 8701 
                 ATGATGGATA CTTTCTCGGC AGGAGCAAGG TGAGATGACA GGAGATCCTG 
               
               
                   
               
               
                 8751 
                 CCCCGGCACT TCGCCCAATA GCAGCCAGTC CCTTCCCGCT TCAGTGACAA 
               
               
                   
               
               
                 8801 
                 CGTCGAGCAC AGCTGCGCAA GGAACGCCCG TCGTGGCCAG CCACGATAGC 
               
               
                   
               
               
                 8851 
                 CGCGCTGCCT CGTCCTGCAG TTCATTCAGG GCACCGGACA GGTCGGTCTT 
               
               
                   
               
               
                 8901 
                 GACAAAAAGA ACCGGGCGCC CCTGCGCTGA CAGCCGGAAC ACGGCGGCAT 
               
               
                   
               
               
                 8951 
                 CAGAGCAGCC GATTGTCTGT TGTGCCCAGT CATAGCCGAA TAGCCTCTCC 
               
               
                   
               
               
                 9001 
                 ACCCAAGCGG CCGGAGAACC TGCGTGCAAT CCATCTTGTT CAATCATGCG 
               
               
                   
               
               
                 9051 
                 AAACGATCCT CATCCTGTCT CTTGATCAGA TCTTGATCCC CTGCGCCATC 
               
               
                   
               
               
                 9101 
                 AGATCCTTGG CGGCAAGAAA GCCATCCAGT TTACTTTGCA GGGCTTCCCA 
               
               
                   
               
               
                 9151 
                 ACCTTACCAG AGGGCGCCCC AGCTGGCAAT TCCGGTTCGC TTGCTGTCCA 
               
               
                   
               
               
                 9201 
                 TAAAACCGCC CAGTCTAGCT ATCGCCATGT AAGCCCACTG CAAGCTACCT 
               
               
                   
               
               
                 9251 
                 GCTTTCTCTT TGCGCTTGCG TTTTCCCTTG TCCAGATAGC CCAGTAGCTG 
               
               
                   
               
               
                 9301 
                 ACATTCATCC GGGGTCAGCA CCGTTTCTGC GGACTGGCTT TCTACGTGTT 
               
               
                   
               
               
                 9351 
                 CCGCTTCCTT TAGCAGCCCT TGCGCCCTGA GTGCTTGCGG CAGCGTGAAG 
               
            
           
         
       
     
     SEQ ID NO:18 
     Nucleotide Sequence of the Expression Vector HCMV-K HuAb-VL1 hum V2 
     (Complete DNA Sequence of a Humanised Light Chain Expression Vector Comprising SEQ ID NO: 13 (humV2=VLm) from 3926-4246) 
     
       
         
           
               
               
               
            
               
                    1 
                 CTAGCTTTTT GCAAAAGCCT AGGCCTCCAA AAAAGCCTCC TCACTACTTC 
                   
               
               
                   
               
               
                   51 
                 TGGAATAGCT CAGAGGCCGA GGCGGCCTCG GCCTCTGCAT AAATAAAAAA 
               
               
                   
               
               
                  101 
                 AATTAGTCAG CCATGGGGCG GAGAATGGGC GGAACTGGGC GGAGTTAGGG 
               
               
                   
               
               
                  151 
                 GCGGGATGGG CGGAGTTAGG GGCGGGACTA TGGTTGCTGA CTAATTGAGA 
               
               
                   
               
               
                  201 
                 TGCATGCTTT GCATACTTCT GCCTGCTGGG GAGCCTGGTT GCTGACTAAT 
               
               
                   
               
               
                  251 
                 TGAGATGCAT GCTTTGCATA CTTCTGCCTG CTGGGGAGCC TGGGGACTTT 
               
               
                   
               
               
                  301 
                 CCACACCCTA ACTGACACAC ATTCCACAGC TGCCTCGCGC GTTTCGGTGA 
               
               
                   
               
               
                  351 
                 TGACGGTGAA AACCTCTGAC ACATGCAGCT CCCGGAGACG GTCACAGCTT 
               
               
                   
               
               
                  401 
                 GTCTGTAAGC GGATGCCGGG AGCAGACAAG CCCGTCAGGG CGCGTCAGCG 
               
               
                   
               
               
                  451 
                 GGTGTTGGCG GGTGTCGGGG CGCAGCCATG ACCCAGTCAC GTAGCGATAG 
               
               
                   
               
               
                  501 
                 CGGAGTGTAT ACTGGCTTAA CTATGCGGCA TCAGAGCAGA TTGTACTGAG 
               
               
                   
               
               
                  551 
                 AGTGCACCAT ATGCGGTGTG AAATACCGCA CAGATGCGTA AGGAGAAAAT 
               
               
                   
               
               
                  601 
                 ACCGCATCAG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG 
               
               
                   
               
               
                  651 
                 GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG 
               
               
                   
               
               
                  701 
                 GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG 
               
               
                   
               
               
                  751 
                 GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC 
               
               
                   
               
               
                  801 
                 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA 
               
               
                   
               
               
                  851 
                 GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG 
               
               
                   
               
               
                  901 
                 GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC 
               
               
                   
               
               
                  951 
                 CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG 
               
               
                   
               
               
                 1001 
                 CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG 
               
               
                   
               
               
                 1051 
                 TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT 
               
               
                   
               
               
                 1101 
                 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC 
               
               
                   
               
               
                 1151 
                 CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT 
               
               
                   
               
               
                 1201 
                 TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGGAC AGTATTTGGT 
               
               
                   
               
               
                 1251 
                 ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC 
               
               
                   
               
               
                 1301 
                 TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA 
               
               
                   
               
               
                 1351 
                 AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC 
               
               
                   
               
               
                 1401 
                 TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT 
               
               
                   
               
               
                 1451 
                 TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT 
               
               
                   
               
               
                 1501 
                 AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT 
               
               
                   
               
               
                 1551 
                 GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT 
               
               
                   
               
               
                 1601 
                 ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA 
               
               
                   
               
               
                 1651 
                 TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC 
               
               
                   
               
               
                 1701 
                 CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG 
               
               
                   
               
               
                 1751 
                 GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA 
               
               
                   
               
               
                 1801 
                 TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG 
               
               
                   
               
               
                 1851 
                 CGCAACGTTG TTGCCATTGC TGCAGGCATC GTGGTGTCAC GCTCGTCGTT 
               
               
                   
               
               
                 1901 
                 TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT 
               
               
                   
               
               
                 1951 
                 GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC 
               
               
                   
               
               
                 2001 
                 GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC 
               
               
                   
               
               
                 2051 
                 ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA 
               
               
                   
               
               
                 2101 
                 CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG 
               
               
                   
               
               
                 2151 
                 AGTTGCTCTT GCCCGGCGTC AACACGGGAT AATACCGCGC CACATAGCAG 
               
               
                   
               
               
                 2201 
                 AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT 
               
               
                   
               
               
                 2251 
                 CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA 
               
               
                   
               
               
                 2301 
                 CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC 
               
               
                   
               
               
                 2351 
                 AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA 
               
               
                   
               
               
                 2401 
                 AATGTTGAAT ACTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT 
               
               
                   
               
               
                 2451 
                 CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA 
               
               
                   
               
               
                 2501 
                 TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG 
               
               
                   
               
               
                 2551 
                 TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC 
               
               
                   
               
               
                 2601 
                 ACGAGGCCCT TTCGTCTTCA AGAATTCAGC TTGGCTGCAG TGAATAATAA 
               
               
                   
               
               
                 2651 
                 AATGTGTGTT TGTCCGAAAT ACGCGTTTTG AGATTTCTGT CGCCGACTAA 
               
               
                   
               
               
                 2701 
                 ATTCATGTCG CGCGATAGTG GTGTTTATCG CCGATAGAGA TGGCGATATT 
               
               
                   
               
               
                 2751 
                 GGAAAAATCG ATATTTGAAA ATATGGCATA TTGAAAATGT CGCCGATGTG 
               
               
                   
               
               
                 2801 
                 AGTTTCTGTG TAACTGATAT CGCCATTTTT CCAAAAGTGA TTTTTGGGCA 
               
               
                   
               
               
                 2851 
                 TACGCGATAT CTGGCGATAG CGCTTATATC GTTTACGGGG GATGGCGATA 
               
               
                   
               
               
                 2901 
                 GACGACTTTG GTGACTTGGG CGATTCTGTG TGTCGCAAAT ATCGCAGTTT 
               
               
                   
               
               
                 2951 
                 CGATATAGGT GACAGACGAT ATGAGGCTAT ATCGCCGATA GAGGCGACAT 
               
               
                   
               
               
                 3001 
                 CAAGCTGGCA CATGGCCAAT GCATATCGAT CTATACATTG AATCAATATT 
               
               
                   
               
               
                 3051 
                 GGCCATTAGC CATATTATTC ATTGGTTATA TAGCATAAAT CAATATTGGC 
               
               
                   
               
               
                 3101 
                 TATTGGCCAT TGCATACGTT GTATCCATAT CATAATATGT ACATTTATAT 
               
               
                   
               
               
                 3151 
                 TGGCTCATGT CCAACATTAC CGCCATGTTG ACATTGATTA TTGACTAGTT 
               
               
                   
               
               
                 3201 
                 ATTAATAGTA ATCAATTACG GGGTCATTAG TTCATAGCCC ATATATGGAG 
               
               
                   
               
               
                 3251 
                 TTCCGCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT GACCGCCCAA 
               
               
                   
               
               
                 3301 
                 CGACCCCCGC CCATTGACGT CAATAATGAC GTATGTTCCC ATAGTAACGC 
               
               
                   
               
               
                 3351 
                 CAATAGGGAC TTTCCATTGA CGTCAATGGG TGGAGTATTT ACGGTAAACT 
               
               
                   
               
               
                 3401 
                 GCCCACTTGG CAGTACATCA AGTGTATCAT ATGCCAAGTA CGCCCCCTAT 
               
               
                   
               
               
                 3451 
                 TGACGTCAAT GACGGTAAAT GGCCCGCCTG GCATTATGCC CAGTACATGA 
               
               
                   
               
               
                 3501 
                 CCTTATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT AGTCATCGCT 
               
               
                   
               
               
                 3551 
                 ATTACCATGG TGATGCGGTT TTGGCAGTAC ATCAATGGGC GTGGATAGCG 
               
               
                   
               
               
                 3601 
                 GTTTGACTCA CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA 
               
               
                   
               
               
                 3651 
                 GTTTGTTTTG GCACCAAAAT CAACGGGACT TTCCAAAATG TCGTAACAAC 
               
               
                   
               
               
                 3701 
                 TCCGCCCCAT TGACGCAAAT GGGCGGTAGG CGTGTACGGT GGGAGGTCTA 
               
               
                   
               
               
                 3751 
                 TATAAGCAGA GCTCGTTTAG TGAACCGTCA GATCGCCTGG AGACGCCATC 
               
               
                   
               
               
                 3801 
                 CACGCTGTTT TGACCTCCAT AGAAGACACC GGGACCGATC CAGCCTCCGC 
               
               
                   
               
               
                 3851 
                 AAGCTTGCCG CCACCATGGA GACCCCCGCC CAGCTGCTGT TCCTGCTGCT 
               
               
                   
               
               
                 3901 
                 GCTGTGGCTG CCCGACACCA CCGGCGACAT TCTGCTGACC CAGTCTCCAG 
               
               
                   
               
               
                 3951 
                 CCACCCTGTC TCTGAGTCCA GGAGAAAGAG CCACTTTCTC CTGCAGGGCC 
               
               
                   
               
               
                 4001 
                 AGTCAGAACA TTGGCACAAG CATACAGTGG TATCAACAAA AAACAAATGG 
               
               
                   
               
               
                 4051 
                 TGCTCCAAGG CTTCTCATAA GGTCTTCTTC TGAGTCTATC TCTGGGATCC 
               
               
                   
               
               
                 4101 
                 CTTCCAGGTT TAGTGGCAGT GGATCAGGGA CAGATTTTAC TCTTACCATC 
               
               
                   
               
               
                 4151 
                 AGCAGTCTGG AGCCTGAAGA TTTTGCAGTG TATTACTGTC AACAAAGTAA 
               
               
                   
               
               
                 4201 
                 TACCTGGCCA TTCACGTTCG GCCAGGGGAC CAAGCTGGAG ATCAAACGTG 
               
               
                   
               
               
                 4251 
                 AGTATTCTAG AAAGATCCTA GAATTCTAAA CTCTGAGGGG GTCGGATGAC 
               
               
                   
               
               
                 4301 
                 GTGGCCATTC TTTGCCTAAA GCATTGAGTT TACTGCAAGG TCAGAAAAGC 
               
               
                   
               
               
                 4351 
                 ATGCAAAGCC CTCAGAATGG CTGCAAAGAG CTCCAACAAA ACAATTTAGA 
               
               
                   
               
               
                 4401 
                 ACTTTATTAA GGAATAGGGG GAAGCTAGGA AGAAACTCAA AACATCAAGA 
               
               
                   
               
               
                 4451 
                 TTTTAAATAC GCTTCTTGGT CTCCTTGCTA TAATTATCTG GGATAAGCAT 
               
               
                   
               
               
                 4501 
                 GCTGTTTTCT GTCTGTCCCT AACATGCCCT GTGATTATCC GCAAACAACA 
               
               
                   
               
               
                 4551 
                 CACCCAAGGG CAGAACTTTG TTACTTAAAC ACCATCCTGT TTGCTTCTTT 
               
               
                   
               
               
                 4601 
                 CCTCAGGAAC TGTGGCTGCA CCATCTGTCT TCATCTTCCC GCCATCTGAT 
               
               
                   
               
               
                 4651 
                 GAGCAGTTGA AATCTGGAAC TGCCTCTGTT GTGTGCCTGC TGAATAACTT 
               
               
                   
               
               
                 4701 
                 CTATCCCAGA GAGGCCAAAG TACAGTGGAA GGTGGATAAC GCCCTCCAAT 
               
               
                   
               
               
                 4751 
                 CGGGTAACTC CCAGGAGAGT GTCACAGAGC AGGACAGCAA GGACAGCACC 
               
               
                   
               
               
                 4801 
                 TACAGCCTCA GCAGCACCCT GACGCTGAGC AAAGCAGACT ACGAGAAACA 
               
               
                   
               
               
                 4851 
                 CAAAGTCTAC GCCTGCGAAG TCACCCATCA GGGCCTGAGC TCGCCCGTCA 
               
               
                   
               
               
                 4901 
                 CAAAGAGCTT CAACAGGGGA GAGTGTTAGA GGGAGAAGTG CCCCCACCTG 
               
               
                   
               
               
                 4951 
                 CTCCTCAGTT CCAGCCTGAC CCCCTCCCAT CCTTTGGCCT CTGACCCTTT 
               
               
                   
               
               
                 5001 
                 TTCCACAGGG GACCTACCCC TATTGCGGTC CTCCAGCTCA TCTTTCACCT 
               
               
                   
               
               
                 5051 
                 CACCCCCCTC CTCCTCCTTG GCTTTAATTA TGCTAATGTT GGAGGAGAAT 
               
               
                   
               
               
                 5101 
                 GAATAAATAA AGTGAATCTT TGCACCTGTG GTTTCTCTCT TTCCTCATTT 
               
               
                   
               
               
                 5151 
                 AATAATTATT ATCTGTTGTT TACCAACTAC TCAATTTCTC TTATAAGGGA 
               
               
                   
               
               
                 5201 
                 CTAAATATGT AGTCATCCTA AGGCGCATAA CCATTTATAA AAATCATCCT 
               
               
                   
               
               
                 5251 
                 TCATTCTATT TTACCCTATC ATCCTCTGCA AGACAGTCCT CCCTCAAACC 
               
               
                   
               
               
                 5301 
                 CACAAGCCTT CTGTCCTCAC AGTCCCCTGG GCCATGGTAG GAGAGACTTG 
               
               
                   
               
               
                 5351 
                 CTTCCTTGTT TTCCCCTCCT CAGCAAGCCC TCATAGTCCT TTTTAAGGGT 
               
               
                   
               
               
                 5401 
                 GACAGGTCTT ACAGTCATAT ATCCTTTGAT TCAATTCCCT GAGAATCAAC 
               
               
                   
               
               
                 5451 
                 CAAAGCAAAT TTTTCAAAAG AAGAAACCTG CTATAAAGAG AATCATTCAT 
               
               
                   
               
               
                 5501 
                 TGCAACATGA TATAAAATAA CAACACAATA AAAGCAATTA AATAAACAAA 
               
               
                   
               
               
                 5551 
                 CAATAGGGAA ATGTTTAAGT TCATCATGGT ACTTAGACTT AATGGAATGT 
               
               
                   
               
               
                 5601 
                 CATGCCTTAT TTACATTTTT AAACAGGTAC TGAGGGACTC CTGTCTGCCA 
               
               
                   
               
               
                 5651 
                 AGGGCCGTAT TGAGTACTTT CCACAACCTA ATTTAATCCA CACTATACTG 
               
               
                   
               
               
                 5701 
                 TGAGATTAAA AACATTCATT AAAATGTTGC AAAGGTTCTA TAAAGCTGAG 
               
               
                   
               
               
                 5751 
                 AGACAAATAT ATTCTATAAC TCAGCAATCC CACTTCTAGA TGACTGAGTG 
               
               
                   
               
               
                 5801 
                 TCCCCACCCA CCAAAAAACT ATGCAAGAAT GTTCAAAGCA GCTTTATTTA 
               
               
                   
               
               
                 5851 
                 CAAAAGCCAA AAATTGGAAA TAGCCCGATT GTCCAACAAT AGAATGAGTT 
               
               
                   
               
               
                 5901 
                 ATTAAACTGT GGTATGTTTA TACATTAGAA TACCCAATGA GGAGAATTAA 
               
               
                   
               
               
                 5951 
                 CAAGCTACAA CTATACCTAC TCACACAGAT GAATCTCATA AAAATAATGT 
               
               
                   
               
               
                 6001 
                 TACATAAGAG AAACTCAATG CAAAAGATAT GTTCTGTATG TTTTCATCCA 
               
               
                   
               
               
                 6051 
                 TATAAAGTTC AAAACCAGGT AAAAATAAAG TTAGAAATTT GGATGGAAAT 
               
               
                   
               
               
                 6101 
                 TACTCTTAGC TGGGGGTGGG CGAGTTAGTG CCTGGGAGAA GACAAGAAGG 
               
               
                   
               
               
                 6151 
                 GGCTTCTGGG GTCTTGGTAA TGTTCTGTTC CTCGTGTGGG GTTGTGCAGT 
               
               
                   
               
               
                 6201 
                 TATGATCTGT GCACTGTTCT GTATACACAT-TATGCTTCAA AATAACTTCA 
               
               
                   
               
               
                 6251 
                 CATAAAGAAC ATCTTATACC CAGTTAATAG ATAGAAGAGG AATAAGTAAT 
               
               
                   
               
               
                 6301 
                 AGGTCAAGAC CACGCAGCTG GTAAGTGGGG GGGCCTGGGA TCAAATAGCT 
               
               
                   
               
               
                 6351 
                 ACCTGCCTAA TCCTGCCCTC TTGAGCCCTG AATGAGTCTG CCTTCCAGGG 
               
               
                   
               
               
                 6401 
                 CTCAAGGTGC TCAACAAAAC AACAGGCCTG CTATTTTCCT GGCATCTGTG 
               
               
                   
               
               
                 6451 
                 CCCTGTTTGG CTAGCTAGGA GCACACATAC ATAGAAATTA AATGAAACAG 
               
               
                   
               
               
                 6501 
                 ACCTTCAGCA AGGGGACAGA GGACAGAATT AACCTTGCCC AGACACTGGA 
               
               
                   
               
               
                 6551 
                 AACCCATGTA TGAACACTCA CATGTTTGGG AAGGGGGAAG GGCACATGTA 
               
               
                   
               
               
                 6601 
                 AATGAGGACT CTTCCTCATT CTATGGGGCA CTCTGGCCCT GCCCCTCTCA 
               
               
                   
               
               
                 6651 
                 GCTACTCATC CATCCAACAC ACCTTTCTAA GTACCTCTCT CTGCCTACAC 
               
               
                   
               
               
                 6701 
                 TCTGAAGGGG TTCAGGAGTA ACTAACACAG CATCCCTTCC CTCAAATGAC 
               
               
                   
               
               
                 6751 
                 TGACAATCCC TTTGTCCTGC TTTGTTTTTC TTTCCAGTCA GTACTGGGAA 
               
               
                   
               
               
                 6801 
                 AGTGGGGAAG GACAGTCATG GAGAAACTAC ATAAGGAAGC ACCTTGCCCT 
               
               
                   
               
               
                 6851 
                 TCTGCCTCTT GAGAATGTTG ATGAGTATCA AATCTTTCAA ACTTTGGAGG 
               
               
                   
               
               
                 6901 
                 TTTGAGTAGG GGTGAGACTC AGTAATGTCC CTTCCAATGA CATGAACTTG 
               
               
                   
               
               
                 6951 
                 CTCACTCATC CCTGGGGGCC AAATTGAACA ATCAAAGGCA GGCATAATCC 
               
               
                   
               
               
                 7001 
                 AGCTATGAAT TCTAGGATCG ATCCAGACAT GATAAGATAC ATTGATGAGT 
               
               
                   
               
               
                 7051 
                 TTGGACAAAC CACAACTAGA ATGCAGTGAA AAAAATGCTT TATTTGTGAA 
               
               
                   
               
               
                 7101 
                 ATTTGTGATG CTATTGCTTT ATTTGTAACC ATTATAAGCT GCAATAAACA 
               
               
                   
               
               
                 7151 
                 AGTTAACAAC AACAATTGCA TTCATTTTAT GTTTCAGGTT CAGGGGGAGG 
               
               
                   
               
               
                 7201 
                 TGTGGGAGGT TTTTTAAAGC AAGTAAAACC TCTACAAATG TGGTATGGCT 
               
               
                   
               
               
                 7251 
                 GATTATGATC TCTAGTCAAG GCACTATACA TCAAATATTC CTTATTAACC 
               
               
                   
               
               
                 7301 
                 CCTTTACAAA TTAAAAAGCT AAAGGTACAC AATTTTTGAG CATAGTTATT 
               
               
                   
               
               
                 7351 
                 AATAGCAGAC ACTCTATGCC TGTGTGGAGT AAGTAAAACC AGTATGTTAT 
               
               
                   
               
               
                 7401 
                 GATTATAACT GTTATGCCTA CTTATAAAGG TTACAGAATA TTTTTCCATA 
               
               
                   
               
               
                 7451 
                 ATTTTCTTGT ATAGCAGTGC AGCTTTTTCC TTTGTGGTGT AAATAGCAAA 
               
               
                   
               
               
                 7501 
                 GCAAGCAAGA GTTCTATTAC TAAACACAGC ATGACTCAAA AAACTTAGCA 
               
               
                   
               
               
                 7551 
                 ATTCTGAAGG AAAGTCCTTG GGGTCTTCTA CCTTTCTCTT CTTTTTTGGA 
               
               
                   
               
               
                 7601 
                 GGAGTAGAAT GTTGAGAGTC AGCAGTAGCC TCATCATCAC TAGATGGCAT 
               
               
                   
               
               
                 7651 
                 TTCTTCTGAG CAAAACAGGT TTTCCTCATT AAAGGCATTC CACCACTGCT 
               
               
                   
               
               
                 7701 
                 CCCATTCATC AGTTCCATAG GTTGGAATCT AAAATACACA AACAATTAGA 
               
               
                   
               
               
                 7751 
                 ATCAGTAGTT TAACACATTA TACACTTAAA AATTTTATAT TTACCTTAGA 
               
               
                   
               
               
                 7801 
                 GCTTTAAATC TCTGTAGGTA GTTTGTCCAA TTATGTCACA CCACAGAAGT 
               
               
                   
               
               
                 7851 
                 AAGGTTCCTT CACAAAGATC CGGGACCAAA GCGGCCATCG TGCCTCCCCA 
               
               
                   
               
               
                 7901 
                 CTCCTGCAGT TCGGGGGCAT GGATGCGCGG ATAGCCGCTG CTGGTTTCCT 
               
               
                   
               
               
                 7951 
                 GGATGCCGAC GGATTTGCAC TGCCGGTAGA ACTCCGCGAG GTCGTCCAGC 
               
               
                   
               
               
                 8001 
                 CTCAGGCAGC AGCTGAACCA ACTCGCGAGG GGATCGAGCC CGGGGTGGGC 
               
               
                   
               
               
                 8051 
                 GAAGAACTCC AGCATGAGAT CCCCGCGCTG GAGGATCATC CAGCCGGCGT 
               
               
                   
               
               
                 8101 
                 CCCGGAAAAC GATTCCGAAG CCCAACCTTT CATAGAAGGC GGCGGTGGAA 
               
               
                   
               
               
                 8151 
                 TCGAAATCTC GTGATGGCAG GTTGGGCGTC GCTTGGTCGG TCATTTCGAA 
               
               
                   
               
               
                 8201 
                 CCCCAGAGTC CCGCTCAGAA GAACTCGTCA AGAAGGCGAT AGAAGGCGAT 
               
               
                   
               
               
                 8251 
                 GCGCTGCGAA TCGGGAGCGG CGATACCGTA AAGCACGAGG AAGCGGTCAG 
               
               
                   
               
               
                 8301 
                 CCCATTCGCC GCCAAGCTCT TCAGCAATAT CACGGGTAGC CAACGCTATG 
               
               
                   
               
               
                 8351 
                 TCCTGATAGC GGTCCGCCAC ACCCAGCCGG CCACAGTCGA TGAATCCAGA 
               
               
                   
               
               
                 8401 
                 AAAGCGGCCA TTTTCCACCA TGATATTCGG CAAGCAGGCA TCGCCATGGG 
               
               
                   
               
               
                 8451 
                 TCACGACGAG ATCCTCGCCG TCGGGCATGC GCGCCTTGAG CCTGGCGAAC 
               
               
                   
               
               
                 8501 
                 AGTTCGGCTG GCGCGAGCCC CTGATGCTCT TCGTCCAGAT CATCCTGATC 
               
               
                   
               
               
                 8551 
                 GACAAGACCG GCTTCCATCC GAGTACGTGC TCGCTCGATG CGATGTTTCG 
               
               
                   
               
               
                 8601 
                 CTTGGTGGTC GAATGGGCAG GTAGCCGGAT CAAGCGTATG CAGCCGCCGC 
               
               
                   
               
               
                 8651 
                 ATTGCATCAG CCATGATGGA TACTTTCTCG GCAGGAGCAA GGTGAGATGA 
               
               
                   
               
               
                 8701 
                 CAGGAGATCC TGCCCCGGCA CTTCGCCCAA TAGCAGCCAG TCCCTTCCCG 
               
               
                   
               
               
                 8751 
                 CTTCAGTGAC AACGTCGAGC ACAGCTGCGC AAGGAACGCC CGTCGTGGCC 
               
               
                   
               
               
                 8801 
                 AGCCACGATA GCCGCGCTGC CTCGTCCTGC AGTTCATTCA GGGCACCGGA 
               
               
                   
               
               
                 8851 
                 CAGGTCGGTC TTGACAAAAA GAACCGGGCG CCCCTGCGCT GACAGCCGGA 
               
               
                   
               
               
                 8901 
                 ACACGGCGGC ATCAGAGCAG CCGATTGTCT GTTGTGCCCA GTCATAGCCG 
               
               
                   
               
               
                 8951 
                 AATAGCCTCT CCACCCAAGC GGCCGGAGAA CCTGCGTGCA ATCCATCTTG 
               
               
                   
               
               
                 9001 
                 TTCAATCATG CGAAACGATC CTCATCCTGT CTCTTGATCA GATCTTGATC 
               
               
                   
               
               
                 9051 
                 CCCTGCGCCA TCAGATCCTT GGCGGCAAGA AAGCCATCCA GTTTACTTTG 
               
               
                   
               
               
                 9101 
                 CAGGGCTTCC CAACCTTACC AGAGGGCGCC CCAGCTGGCA ATTCCGGTTC 
               
               
                   
               
               
                 9151 
                 GCTTGCTGTC CATAAAACCG CCCAGTCTAG CTATCGCCAT GTAAGCCCAC 
               
               
                   
               
               
                 9201 
                 TGCAAGCTAC CTGCTTTCTC TTTGCGCTTG CGTTTTCCCT TGTCCAGATA 
               
               
                   
               
               
                 9251 
                 GCCCAGTAGC TGACATTCAT CCGGGGTCAG CACCGTTTCT GCGGACTGGC 
               
               
                   
               
               
                 9301 
                 TTTCTACGTG TTCCGCTTCC TTTAGCAGCC CTTGCGCCCT GAGTGCTTGC 
               
               
                   
               
               
                 9351 
                 GGCAGCGTGA AG 
               
            
           
         
       
     
     Example 9 
     In Vitro Efficacy of CD45RO/RB Binding Humanised Antibodies 
     To determine the efficacy of the CD45R0/RB binding humanised antibodies VHE/humV1 and VHQ/humV1 in comparison to the chimeric antibody the ability to induce apoptosis in human T cells and also the ability to inhibit human T cell proliferation is analysed. 
     Cells and Reagents 
     Peripheral blood mononuclear cells (PBMC) are isolated from leukopheresis samples of healthy human donors with known blood type, but unknown HLA type by centrifugation over Ficoll-Hypaque (Pharmacia LKB). PBMC used as stimulators are first depleted of T and NK cells by using CD3-coated ferromagnetic beads (Miltenyi). Beads and contaminating cells are removed by magnetic field. T cell-depleted PBMC are used as stimulator cells after irradiation (50 Gy). CD4 +  T cells are used as responder cells in MLR and are isolated from PBMC with a CD4 T cell negative selection kit (Miltenyi). 
     The obtained cells are analyzed by FACScan or FACSCalibur (Becton Dickinson &amp; Co., CA) and the purity of the obtained cells is &gt;75%. Cells are suspended in RPMI1640 medium supplemented with 10% heat-inactivated FCS, penicillin, streptomycin and L-glutamine. 
     Apoptosis Assays 
     Human PBMC of three healthy voluntary donors are cultured in growth medium (RPMI1640+10% FCS) overnight (&lt;16 h) in the presence of CD45R0/RB binding chimeric mAb, humanized antibodies (VHE/humV1 and VHQ/humV1) or anti-LPS control mAb. If indicated, a cross-linking reagent, F(ab′) 2 -fragment of goat anti-human IgG (Cat. No. 109-006-098, JacksonLab) is included at a μg/ml concentration being twice as high as the sample&#39;s anti-CD45 antibodies concentration. The PBS-concentration in all wells introduced by the antibody reagents is kept constant among all samples, namely at 20% (v/v) for samples without cross-linker or at 40% (v/v) for samples with cross-linker. Earlier experiments demonstrate that the amount of PBS does not affect the readout. 
     After overnight culture in the presence of the antibodies, the samples are subjected to flow cytometry analyses and stained with the apoptosis marker AnnexinV-FITC (Cat. No. 556419, BD/Pharmingen) and the T cell marker CD2-PE (Cat. No. 556609, BD/Pharmingen). The samples are run in a Becton Dickinson FACSCalibur instrument and the data are analyzed using the CellQuest Pro Software. 
     From the data collected, curves are fitted using the software Origin v7.0300 The equation used for fitting is 
     
       
         
         
             
             
         
       
     
     A 1 : final value (for fitting sessions set to “shared” and “floating”)
 
A 2 : initial value (for fitting sessions set to “shared” and “floating”)
 
p: power
 
X 0 : ED 50 ; IC 50  (see below).
 
     In the absence of cross-linker, VHE/humV1 is most effective, with an ED 50  value of 148±71 nM, followed by VHQ/humV1 with 377±219 nM. CD45R0/RB binding chimeric antibody is less effective with an ED 50  value of 2440±1205 nM. 
     In the presence of a cross-linking antiserum, the ED 50  values are shifted dramatically towards higher efficacy by at least two orders of magnitude. In addition, the presence of cross-linker permitted higher levels of apoptosis at very high antibody concentrations, now reaching up to 80%, whereas the absence of cross-linker only allowed for up to 50% of apoptosis. In the presence of cross-linker, the curves (antibody concentration/% apoptosis) are bi-modal with two plateaus: the first plateau is reached at low antibody concentrations (˜5 nM), where the apoptosis level corresponds to the maximum level obtained in the absence of cross-linker. The second plateau is reached at high antibody concentrations (˜500 nM) and apoptosis is observed within 70-80% of the T cell population. 
     Both CD45R0/RB binding humanised mAb are equally effective and better or equal compared to CD45R0/RB binding chimeric mAb with respect to their ability to induce apoptosis in primary human T cells. 
     Mixed Lymphocyte Reaction Assays 
     One×10 3  PBMC or 5×10 4  of CD4 +  cells are mixed with 1×10 5  or 5×10 4  T cells-depleted irradiated (50 Gy) PBMC in each well of 96-well culture plates in the presence or absence of the different concentrations of mAb. 
     The mixed cells are cultured for 5 days and proliferation is determined by pulsing the cells with  3 H-thymidine for the last 16-20 hours of culture. MLR inhibition at each antibody concentration is expressed as percentage inhibition as described in Example 2. 
     The effect of increasing concentrations of VHE/humV1 and VHQ/humV1 on MLR is evaluated in three responder:stimulator combinations. All antibodies inhibit the MLR in a dose-dependent manner. The IC 50  values (see above) are similar for the humanized Ab VHE/humV1 (7±7 nM) and VHQ/humV1 (39±54 nM). Both humanised antibodies are more potent in inhibiting MLR than the parental chimeric antibody (IC 50  of 347±434 nM). As usually seen with MLR experiments, donor variability is high in these experiments. 
     Example 10 
     Specificity of CD45RB/RO Binding Molecule 
     The CD45 molecule is expressed on all leukocytes. However, different CD45 isoforms are expressed by the various leukocyte subsets. In order to determine the leukocyte subset reactivity of CD45RB/RO binding chimeric antibody molecule immunofluorescent labeling of human leukocytes with subset-specific markers and simultaneous immunofluorescent labeling with a dye-conjugated CD45RB/RO binding chimeric antibody is performed, followed by flow cytometry analysis. Briefly, specific subsets of a freshly isolated preparation of human peripheral blood mononuclear cells (PBMC), human platelets, human peripheral blood neutrophils or human bone-marrow derived hematopoietic stem cells are identified by incubation with phycoerythrin-coupled antibodies against CD2 (T lymphocytes), CD14 (monocytes), CD19 (B lymphocytes), CD34 (stem cells), CD42a (platelets), CD56 (natural killer cells) or CD66b (granulocytes). Simultaneous binding of a FITC-labeled chimeric CD45RB/RO binding molecule is detected on T lymphocytes, monocytes, stem cells, natural killer cells and granulocytes, but not on platelets or B lymphocytes. 
     Example 11 
     In Vitro Induction of Suppressor T Cells (T Regulatory Cells) and of Functionally Paralyzed T Cells 
     To demonstrate the ability of a CD45RO/RB binding chimeric antibody to induce suppressor T cells, the antibody is included at various concentrations during the generation of CD8+ T cell lines reactive with the antigen matrix protein 1 (MP1) of  hemophilus influenza . These lines are generated through repeated co-culture of CD8+ human lymphocytes with CD14+ human monocytes pulsed with the antigen. Later on, CD14+ monocytes can be replaced with a human leukocyte antigen-2 positive cell line as an MP1 antigen-presenting cell (APC). If such MP1-specific CD8+ T cells from a culture including CD45RO/RB binding chimeric antibody are mixed with freshly isolated human CD8+ T cells and this mixture of cells is stimulated with the MP1 antigen on APC, the addition of CD8+ T cells from the culture in the presence of CD45RO/RB binding molecule is able to reduce the IFN-γ production in an antibody-dose-dependent fashion. No CD45RO/RB binding chimeric antibody is present during this IFN-γ assay culture, indicating that the pre-treatment with the CD45RO/RB mAb has induced CD8+ T cells capable of suppressing the activation of freshly isolated T cells. Because of this induction of suppressor T regulatory cells by the CD45RO/RB binding chimeric antibody, the antibody may be useful in diseases, where a dysregulated and/or activated T cell population is thought to contribute to the pathology. Examples of such diseases include autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease and allergies. 
     To demonstrate the ability of a chimeric CD45RO/RB binding molecule to render T cells hyporesponsive (anergic) to further stimulation, i.e. to functionally paralyze T cells, the antibody is included during the generation of CD8+ T cell lines reactive with the antigen matrix protein 1 (MP1) of  hemophilus influenza  as outlined above. Paralysis is assessed by activating the T cells (exposed prior to CD45RO/RB binding chimeric antibody) with MP1 antigen presented by APC. No CD45RO/RB binding molecule is present in this culture. CD8+ T cells not exposed to CD45RO/RB binding chimeric antibody previously produce IFN-γ upon the mentioned stimulus. In contrast, CD8+ T cells pre-treated with CD45RO/RB binding chimeric antibody show a markedly reduced to inexistent production of this cytokine in response to the antigen-stimulus, demonstrating the CD45RO/RB binding chimeric antibody&#39;s ability to functionally paralyze human T cells. Because of this induction of functional T cell hyporesponsiveness by the CD45RO/RB binding molecule, the antibody may be used in diseases, such autoimmune diseases, transplant rejection, psoriasis, inflammatory bowel disease or allergies, where an activated T cell population is thought to contribute to the pathology. 
     Example 12 
     In Vivo Studies in SCID-hu Skin Mice 
     In this study, the utility of the CD45RB/RO binding chimeric antibody in a Psoriasis model system is tested. Human skin from normal individuals is transplanted to SCID (SCID-hu Skin) mice and the inflammatory process is mimicked by transferring mononuclear cells of unrelated donors into the SCID-hu Skin mice. 
     Transplantation of Human Adult Skin in SCID Mice (SCID-hu Skin Mice) 
     Two small pieces (1 cm 2 ) of human adult skin (obtained from the West Hungarian Regional Tissue Bank; WHRTB, Gyor) consisting of the entire epidermis, the papillary dermis and part of the reticular dermis, are transplanted at the right and left upper-back sides of SCID mice C.B 17/GbmsTac-Prkdc scid  Lyst bg  mice (Taconic, Germantown, N.Y.) in replacement of mouse skin. The quality of the grafts is monitored during 5-6 weeks following transplantation and successfully transplanted mice (SCID-hu Skin mice, generally &gt;85%) are selected for in vivo testing of CD45RB/RO binding chimeric antibody. 
     Engraftment of Human Mononuclear Cells in SCID Mice 
     Mononuclear splenocytes (Spl) are isolated from human adult spleen biopsies (WHRTB, Gyor) after cell suspension (using a cell dissociation sieve equipped with a size 50 mesh) and standard density gradient procedures. Aliquots of ˜5×10 8  Spl are re-suspended in 1.5 ml of RPMI-10% FCS and injected intraperitoneally (i.p.), on experimental day 0, into the SCID-hu Skin mice. These Spl numbers have been found in previous experiments to be sufficient to induce a lethal xeno-GvHD in &gt;90% of the mice within 4-6 weeks after cell transfer. 
     Antibody Treatment of SCID-hu Skin Mice 
     SCID-hu Skin mice, reconstituted with human Spl, are treated with CD45RB/RO binding chimeric antibody or with anti-LPS control mAb at day 0, immediately after mononuclear cell injection, at days 3 and 7 and at weekly intervals thereafter. Antibodies are delivered subcutaneously (s.c.) in 100 μl PBS at a final concentration of 1 mg/kg body weight (b.w.). 
     Evaluation of Anti-CD45 Treatment 
     The efficacy of CD45RB/RO binding chimeric antibody is assessed by the survival of the transplanted mice and by monitoring the rejection of the skin grafts. The significance of the results is evaluated by the statistical method of survival analysis using the Log-rank test (Mantel method) with the help of Systat v10 software. At the end of the experiment biopsies of human skin grafts and mouse liver, lung, kidney and spleen are obtained from sacrificed mice for histological purposes. All mice are weighed at the beginning (before cell transfer) and throughout the experiment (every two days) as an indirect estimation of their health status. Linear regression lines are generated using the body weight versus days post-PBMC transfer values obtained from each mouse and subsequently, their slopes (control versus anti-CD45 treated mice) are compared using the non parametric Mann-Whitney test. 
     Results 
     The human skin grafts are very well tolerated by the SCID mice. Initially, the grafts undergo a period of keratinocyte hyperproliferation resulting in the formation of hyperkeratotic crusts. About 5 weeks after transplantation, the crusts fall off the grafts and reveal a tissue containing all the characteristic structures observed in normal human skin. During this process, the human skin grafts fuse with the adjacent mouse skin and generate a network of freshly grown human vessels that connect the grafts with the underlying mouse tissue. The circulating human Spl transferred into SCID-hu Skin mice (at experimental day 0, approx. 6 weeks after skin transplantation) infiltrate the skin grafts and after recognition of alloantigen molecules expressed on the human skin mount an inflammatory response that in some cases completely destroy the graft. 
     Treatment of these mice with CD45RB/RO binding chimeric antibody suppresses the inflammatory process and prevents the rejection of the human skin grafts. In contrast, the sample obtained from the control treated mouse shows a massive infiltration with multiple signs of necrosis and a dramatic destruction of the epidermis. This process is easily monitored by eye and documented by simple photography of the mice. 
     Six out of six SCID-hu Skin mice transferred with allogeneic human Spl and treated with control anti-LPS mAb show a strong inflammatory response clearly visible by eye 23 days after mononuclear cell transfer. All mice show considerable lesions, including erythema, scaling and pronounced pustules. In contrast the skin grafts of all mice treated with CD45RB/RO binding chimeric antibody have a normal appearance. The dramatic differences between the two groups of mice is specifically due to the antibody treatment since the human skin of all mice have an identical look at the beginning of the experiment. This aspect is not changed until the second week after cell transfer, the time at which the control group started to developed skin lesions. The experiment is terminated at day 34 after mononuclear cell transfer. By that time, one of the control mice is already dead (day 30) and four other are sacrificed (days 27, 27, 27 and 30) due to a strong xeno-GvHD. The pathologic reactions observed in the antibody control treated mice also correlates with a loss of body weight in these animals. 
     In contrast, the CD45RB/RO binding chimeric antibody treated group displays a healthy status during the whole experimentation time.