SOLUTION BASED TWO-DIMENSIONAL SEPARATION AND DETECTION OF AMPHOTERIC SUBSTANCES

Abstract: A method and apparatus for separating mixtures of amphoteric substances uses two dimensions of separation. A sample valve (8, 9) is used to introduce a sample into the mobile phase (7b) for entry into the first dimension of separation, typically a chromatography column (10). The separated sample is passed to the second dimension, which is formed by a series of stages (11, 18, 19). The first stage (11) has a manifold having an anode (13), cathode (12) and a membrane (15) of a specific pK, defining two channels (16a, 16b). The amphoteric molecules are segregated between the two channels and the segregated molecules are passed to a second stage, where each channel (16a, 16b) is further split into two channels (16e, 16f and 16c, 16d), respectively. In each subsequent stage (19), the number of channels is doubled from the previous stage, increasing the degree of separation by pH.

Description: SOLUTION BASED TWO-DIMENSIONAL SEPARATION AND DETECTION OF AMPHOTERIC SUBSTANCES TECHNICAL FIELD AND INDUSTRIAL APPLICABILITY OF INVENTION This invention relates to a method and apparatus to separate and detect amphoteric substances, such as proteins, in complex mixtures in two or more dimensions such that all component analytes are segregated to a predetermined level of resolution in multiple channels. BACKGROUND OF THE INVENTION Two-dimensional separation of amphoteric molecules, such as proteins, is an established art. One of the most common embodiments of this art is 2D-gel electrophoresis, first introduced by P.H. O'Farrell in "High-Resolution Two- dimensional Electrophoresis of Proteins" Journal of Biological Chemistry, 250: 4007- 4021 (1975). This art involves the separation of proteins embedded in hydrophilic gels, such as acrylamide, using two physio-chemical parameters. Amphoteric molecules (or larger structures such as cellular organelles or even whole cells) are defined by a plurality of positive and negative functional chemical groups located on an accessible surface that can be titrated or buffered by changes in the pH of the solution environment in which they find themselves. Amphoteric entities lose negative and/or gain positive charges in a more acidic environment. Conversely, amphoteric entities lose positive and/or gain negative charges in a more basic environment. When placed in an externally applied direct current electrical field, within a pH gradient environment, in which the acidic pH region is aligned with the anode and the basic region with the cathode, amphoteric entities will migrate, based on their net charge, towards one of the electrodes. As they do so, they will gain or lose charges until they reach a pH condition at which they have no net charge. This pH condition is known as the isoelectric point and the process is known as isoelectric focusing. If the amphoteric entities diffuse or drift away from this pH condition they will again migrate so as to return to the pH condition of their isoelectric point. In 2D gel electrophoresis, separation is provided first according to isoelectric point in a contiguous pH gradient wherein molecules migrate, as described, in an applied electrical field to the position in the gradient where the net charge on a molecule is zero (i.e. isoelectric focusing). The second physio-chemical parameter involves rapid denaturation of the isoelectrically focused molecules while still embedded in the gel matrix. This denaturation is provided by rapidly diffusing a detergent, typically sodium dodecyl sulfate (SDS) into the gel matrix, such that all amphoteric molecules are coated with an overwhelming negative charge before they can diffuse away from their isoelectric position in the gel, thus rendering them non-amphoteric. The SDS treated isoelectric focusing acrylamide matrix, typically in the shape of a tube or strip, is quickly brought in contact with a second acrylamide gel matrix, typically in the shape of a slab, by application along one side. An electrical potential is applied to the second acrylamide gel perpendicular to the isoelectric focusing gel strip so as to transfer and further separate the denatured molecules. With the native charges of the molecule masked by a constant amount of negatively charged SDS per weight of protein, the molecules move through the second gel matrix according to their molecular weight alone, towards the anode. The result is a two-dimensional map in which each protein migrates to an embedded spot in the second dimension gel that corresponds to its molecular weight and isoelectric point. This prior art provides a means of segregating many thousands of individual analytes in a complex mixture of proteins, such as a cell lysate. The resolving power of 2D-gel electrophoresis is unmatched by other current methods of protein separation and is well optimized with recent advances. The book by P.G. Righetti 2 Immobilized pH Gradients: Theory and Methodology Elsevier: New York (1990) provides a detailed description of the art of 2D-gel electrophoresis using an improved method of pH gradient formation called immobilized pH gradients. There are, however, a number of disadvantages to the art of 2D-gel electrophoresis as it is currently practiced, including the improved method of pH gradient formation. These disadvantages stem from the nature of the separation method wherein the sample is embedded in the separating gel matrix at the end of the analysis. For example, the technique of 2D-gel electrophoresis is difficult to perform in a reproducible manner (see Corbett et al., Positional reproducibility of protein spots in two-dimensional polyacrylamide gel electrophoresis using immobilized pH gradient isoelectric focusing in the first dimension: an inter-laboratory comparison, Electrophoresis 1994, 15 1205-1211) and requires much manual manipulation in equilibrating and transferring the first dimension isoelectric focusing strip or tube gel onto the second dimension gel within a narrow time span. Since the separated proteins are embedded in the separating matrix at the end of the process, the available detection methods are limited to those methods that provide a captured image in two dimensions corresponding to the separation pattern of the proteins. These methods include visible or fluorescent staining of the proteins or scanning measurements of radioactivity. Such imaging methods are generally of lower sensitivity than solution based detection methods such as on-line flow cell analysis. An additional disadvantage of the current art of 2D-gel electrophoresis is that the proteins are completely denatured and devoid of most if not all biological activity. Also, in the current art of 2D-gel electrophoresis, if it is desired to further characterize the embedded analytes such as proteins, they must be electro-transferred from the entire gel onto a binding membrane or punched out of the gel as individual spots and eluted. Such methods are time consuming, introduce variability and contaminants in any subsequent analysis and are limited to providing analysis of characteristics that do not depend on biological activity. These disadvantages, found in all versions of the current art, can be overcome if a method were devised that combined the high resolution, near baseline separation of many thousands of individual proteins, as found in existing 2D-gel methods, with a system that maintained the analyte molecules in free solution in a continuously flowing mobile phase, instead of embedded in the separation matrix. Such a method would allow for major improvements in detection sensitivity, ease of use and potential recovery of non-denatured proteins for further analysis. The current invention achieves this goal of an on-line solution based multidimensional separation system. Furthermore, if the separation or detection methodology in this new method included the same two physio-chemical parameters as 2D-gel electrophoresis, i.e. isoelectric point and molecular weight, then a large body of existing archived data could be more easily correlated to any subsequent data. Under these conditions, the data derived from the current invention can be correlated to the large body of existing archived 2D gel data using computational reconstruction. The general method of separation of molecules using various septa or membranes to divide an analyte stream into a series of parallel laminar flow channels with a typically transverse applied electrical field in order to transfer these molecules between channels is well established. These techniques, in general, provide for a non-embedded sample. Examples of such apparatus are described in U.S. Patent No. 4,204,929 (Bier), U.S. Patent No. 4,362,612 (Bier), U.S. Patent No. 4,963,236 (Rodkey et al.) and its continuation in part U.S. Patent No. 5,160,594 (Huff et al.), U.S. Patent No. 4,971 ,670 (Faupel et al.), U.S. Patent No. 5,540,826 (Bier) and many others. In general, and in contrast to the current invention, these methods of separation make no attempt to optimize the width or geometry of the channels containing the analyte molecules in order to achieve rapid separation equilibrium. The typical widths of the laminar channels and the distances between the anode and the cathode are substantial. For instance, the IsoPrime™ multi-compartment electrofocusing unit (PI 8) manufactured by Hoefer Scientific Instruments, 654 Minnesota St., San Francisco, CA and based on US Patent No. 4,971 ,670 (Faupel et al.), provides a minimum of three laminar channels with a minimum volume of 8 mis within each chamber. The ratio of separating membrane surface area to volume is 1 cm 2 / ml or less. Instead of rapid equilibrium, these existing separation methods typically utilize strategies of gradual equilibrium with re-circulation in which the sample is recycled through the separating apparatus in many passes obtaining partial separation with each pass. This is sometimes referred to as interative purification. The use of a dividing membrane or septum in these general methods and a buffering membrane in particular is central to the current invention. A prior art has been established in U.S. Patent No. 4,971 ,670 (Faupel et al.), for the preparative separation of proteins and other amphoteric molecules in which the entire analyte stream is recycled many times through a few widely spaced channels (each typically connected to a re-circulating pump) between specific buffering membranes in a cylindrical apparatus. These buffering membranes have an immobilized aggregate pK value titrated during manufacture to provide a set pH within each membrane. When a transverse electrical field is applied, amphoteric analyte molecules will move through such membranes towards the electrode having the opposite charge from the charge on the amphoteric molecule. This electrophoretic movement will continue until the buffering pH of an incident membrane modifies the charge on the amphoteric molecule such that it no longer has an opposite charge from the charge of electrode to which it had been moving. The amphoteric molecule will then remain in one of the laminar spaces between membranes. The method recycles the sample through each channel many times and achieves gradual equilibrium over time scales of hours with a single stage of separation. In all the aforementioned methods in which a transverse electrical field is applied to a series of parallel flow channels, any given analyte molecule, if randomly located in a laminar space far removed from the laminar space containing its isoelectric point, must travel a considerable distance through two or more membranes and perhaps through all the membranes by electromotive force. This prior art using re-circulating analyte streams is well suited for the purification of proteins on a preparative scale as described in articles such as Righetti et al., PrlME Purification of Human Monoclonal Antibodies Against HIV gp-41 Hoefer IsoPrime Application note No.2, Hoefer Scientific Instruments (1993) where one or at most a few analytes are to be segregated in laminar spaces typically with very narrow pH boundaries chosen to include the isoelectric points of the pre-determined analyte or analytes. Another example of such an application is the final stage "polishing" purification of a recombinant protein for commercial use as a biological pharmaceutical, in which minor contaminates or degradation products are removed by their differing isoelectric points from the main product described in Righetti et al., PrlME Purification of Recombinant Human Growth Hormone Hoefer IsoPrime Application note No.3, Hoefer Scientific Instruments (1993) This prior art is not, however, well suited to the analytical segregation of a complex mixture of amphoteric molecules of unknown isoelectric point, such as a protein cell lysate which may contain more than 10,000 separate analytes. For complex mixture analysis in which perhaps, by example, 2048 discrete channels of segregation are used, a single stage apparatus of the type described in U.S. Patent No. 4,971 ,670 (Faupel et al.) or other prior art is impractical. Such a single stage apparatus would require 2047 membranes and would require an almost impossible number of membrane traverses for some of the analyte molecules that happened to be randomly located on the other end of the apparatus at the onset. By contrast, in the current invention the topographic arrangement and geometry are optimized to provide the minimum transfer distance and minimum number of membrane traverses for all the analytes in a complex mixture. The maximum number of membrane traverses for any analyte molecule using the current invention is equal to the number of stages of segregation, which for the example of 2048 discrete channels is 11 , most analyte molecules will make fewer membrane traverses and some analyte molecules will, by chance, make none. The current invention differs from the prior art additionally in the manner in which coulombic heating is dissipated. In the prior art, U.S. Patent No. 5,160,594(Huff et al.) for instance, analyte streams are typically continuously and separately re-circulated into and out of each separation channel by hydraulic means while its composition slowly changes by lateral movement of analytes through the membranes. In this manner, the analyte stream itself provides heat dissipation by means of radiative or active external cooling during the time it is not within the electrophoretic field. By contrast, in the current invention, the surface area of each buffering membrane and electrode is very large relative to the volume of the individual channels (typically 100 cm 2 / ml) and the distance that heat must flow to exit an individual channel is very short. In the current invention, there is only a single separating membrane between each pair of electrodes, and the analyte stream reaches isoelectric equilibrium across this buffering membrane within a single pass through the particular stage without recirculation, as in the prior art. In addition each electrode is provided with a separate cooling fluid flow to dissipate coulombic heat and to remove small molecules such as salts. The advantage of removing salts in isoelectric focusing is well established. For instance, In U.S. Patent No. 4,396,447(Jain et al.) a prior stage of electrodialysis was introduced to the analyte stream before isoelectric focusing. In the current invention the removal of low molecular weight ionic substances from the analyte stream is simultaneous with isoelectric segregation. These and other differences in the current invention (combined with a first dimensional separation method) provide a novel means for the on-line segregation of amphoteric molecules that cannot be attained by the prior art. SUMMARY OF THE INVENTION The invention comprises a method and an apparatus for the separation of amphoteric substances, such as proteins in two dimensions in a continuously flowing mobile phase in which the outflow is segregated into as many pH range sub-fractions as is desired. Complex mixtures of soluble amphoteric substances, such as the proteins found in total or partial cell lysates, represent a preferred sample to be analyzed by the present invention. Said samples are applied to a first dimension separation means using a sample injection apparatus of the type typically used for sample application in chromatographic workstations. These said sample injection apparatus are of the type having a sample loop or chamber pre-filled with the analyte sample such that it can be incorporated into the mobile phase stream by mechanical means. " In general, a first dimension separation method is provided that can partition the various analytes between a stationary phase matrix and a liquid mobile phase using some physio-chemical parameter that varies among the various analytes. This provides a continuously outflowing stream in which the various analytes are distributed in retention times within the apparatus, thus having differential elution times. In one preferred embodiment of the present invention, the first dimension method for separation of the complex mixture is high-resolution ion exchange chromatography. This separation method can include strong ion exchange matrices that can operate at extremes of pH such quaternary ammonium or sulfo-propyl ion exchange or weak ion exchange matrices such as diethylaminoethyl or carboxymethyl ion exchange. In another preferred embodiment of the present invention, the first dimension method for separation is high-resolution size exclusion chromatography, wherein the fractional volume of a stationary phase matrix accessible to a particular molecule is inversely related to the Stokes radius of said molecule that, in turn, is directly related to the molecular weight of said molecule. In this preferred embodiment, the first dimension size exclusion separation method provides for a continuously outflowing mobile phase containing soluble analytes of comparable molecular size and molecular weight at any given moment. In all preferred embodiments, the first dimensional separation provides a means of high resolution partitioning of the analyte molecules to achieve a maximum variation in elution times of said analyte molecules. A further means is provided to introduce the partitioned outflowing mobile phase of the first dimensional separation means into a second dimensional segregating apparatus wherein a serial and parallel arrangement of a plurality of laminar manifolds is provided in a plurality of stages such that the continuously flowing mobile phase is subdivided into two discrete isoelectric point (pH) ranges of amphoteric molecules in two channels within each stage. The outflow from each said channel is connected to the inlets of two or more channels of a subsequent stage manifold. This configuration provides for further binary separations, subdividing the first two discrete isoelectric point pH ranges into four pH ranges. Further stages generate a "power of two" increase in the number of ever narrower isoelectric point pH ranges (i.e. 8 pH ranges, 16, 32, 64 etc.) It is a desired feature of the present invention that the residence time of the mobile phase within each said stage is comparable in order to reach equilibrium under equivalent conditions. With the overall flow rate of the apparatus constant for all separation dimensions and stages, a preferred embodiment of the current invention provides for a concomitant "power of two" decrease in the number or volume of laminar channels per stage dedicated to a particular pH segregation (see table 1 ). This preferred embodiment has the advantage that as the isoelectric point range narrows with each stage and the approximate number of species of amphoteric molecules within each range decreases concomitantly; the apparatus provides roughly equal amounts of analyte per unit of laminar manifold capacity. In one preferred embodiment where the first dimension is high-resolution size exclusion chromatography, for example, the final pH segregation stage will consist of 2 n pH range segregation membranes (wherein n= the number of stages) and 2 (n+1) outflowing streams of amphoteric molecules, such as proteins. In this particular embodiment, the analyte molecules will be distributed by decreasing molecular weight with time and by isoelectric point in discrete pH range channels. In the other preferred embodiments of the first dimensional separating means, the outflowing streams of amphoteric molecules will be distributed by isoelectric point in discrete pH range channels and one other physio-chemical separating parameter. A means is provided to detect and collect said analyte molecules from all the multiple discrete pH range channels simultaneously. Said means of detection can operate continuously at the time of outflow from the second dimension separation means. It can also be delayed in time or rate of detection from the rate of outflow of the analyte stream. Said means of detection includes all of the established methods of flow cell detection or immobilized sample detection including but not limited to; optical density detection, dynamic and static fluorescence detection, conductivity detection, electrochemical detection, dynamic and static light scattering detection and mass spectroscopic detection. Following analyte detection in each of the outflowing channels, the data can be plotted to create a minimum of a two-dimensional map of the analytes. In the preferred cell lysate sample, where the amphoteric molecules are proteins, this two- dimensional map can be comparable to the two-dimensional maps of 2D-gel electrophoresis except that the samples are not embedded in a matrix. TABLE 1 is a representative example of a preferred arrangement of stages and the replicate number of manifolds for each stage resulting in a 2048 channel pH separation. Stage # pH # Replicate # Outflow Total Σ total n membrane manifold plates pH ranges manifold plates manifold plates types /stage /stage /stage / stage by stage 0 1 32 2 32 32 1 2 16 4 32 64 2 4 8 8 32 96 3 8 4 16 32 128 4 16 2 32 32 160 5 32 1 64 32 192 6 64 1 / 2 128 32 224 7 128 256 32 256 8 256 1/8 512 32 288 9 512 1/16 1024 32 320 10 1024 1/32 2048 32 352 BRIEF DESCRIPTION OF THE DRAWINGS Comprehension of the invention is facilitated by reading the following detailed description in conjunction with the annexed drawings in which: FIG. 1 is a schematic representation of the various components of the method showing the overall relationship between the major components and the direction of flow of the continuous stream of analytes wherein the first dimension solution based separation method maintains a non-varying (isocratic) mobile phase composition. FIG. 2 is a schematic representation of the various components of the method showing the overall relationship between the major components and the direction of flow of the continuous stream of analytes wherein the first dimension solution based separation method uses a time variant (gradient) mobile phase composition. FIG. 3A is a schematic cross sectional representation of a preferred arrangement of a single cathode plate assembly showing channel and fluid connections. FIG. 3B is a schematic vertical cross section through one typical cathode plate showing the relative position in the vertical dimension of transverse channels for the collection of outflowing analyte streams. FIG. 3C is a schematic cross sectional view cut vertically and horizontally through a single manifold assembly showing the outflowing cathodic analyte stream bypassing an end joint through a short transverse channel and emptying into a pooling cathodic analyte stream transverse channel. FIG. 4 is a schematic cross sectional representation of a preferred arrangement of a single anode plate assembly showing channel and fluid connections. FIG. 5 is a schematic cross sectional representation of a preferred arrangement of a single semi-permeable buffering membrane support assembly showing a plurality of semi-permeable buffering membranes and fluid connections. FIG. 6A is a schematic cross sectional representation of a preferred arrangement of a single spacer gasket used to form and maintain the laminar analyte flow channels on the anodic side of a buffering membrane wherein said channels are in the higher of two positions as aligned with channels formed into said anode. FIG. 6B is a schematic cross sectional representation of a preferred arrangement of a single spacer gasket used to form and maintain the laminar analyte flow channels on the cathodic side of a buffering membrane wherein said channels are in a lower of two positions as aligned with channels formed into said cathode. FIG. 7A is a schematic cross sectional representation of a preferred arrangement of a single spacer gasket used to form and maintain the laminar analyte flow channels on the anodic side of a buffering membrane wherein said channels are in the lower of two positions as aligned with channels formed into said anode. FIG. 7B is a schematic cross sectional representation of a preferred arrangement of a single spacer gasket used to form and maintain the laminar analyte flow channels on the cathodic side of a buffering membrane wherein said channels are in the higher of two positions as aligned with channels formed into said cathode. FIG. 8 is a cross sectional representation of a preferred arrangement of a single semi-permeable buffering membrane manifold assembly; comprised of a cathode plate assembly (FIG. 3), a low molecular weight cutoff semi-permeable membrane (13b), a cathode side spacer gasket (FIG. 7B), a semi-permeable buffering membrane assembly (FIG. 5), an anode side spacer gasket (FIG. 6A), a second low molecular weight cutoff semi-permeable membrane (13a) and an anode plate assembly (FIG. 4). A single manifold assembly shares a cathode plate on one side and an anode plate on the other side with adjacent manifold assemblies. FIG. 9 is a schematic representation of a preferred arrangement of a further assembly of individual manifold assemblies (11) into two representative stages of binary segregating manifolds; wherein individual manifold assemblies are overlapped between stages to provide connecting channels between stages. FIG. 10 is a schematic representation of a preferred arrangement of the overall assembly of individual binary pH segregating stages into a continuous flow multichannel second dimension separating means comprised of a inflow pressure cap, a plurality of individual separating stages, a outflow pressure cap, a means of simultaneous collection of fractions from each channel and a means of immobilizing part of the analyte stream for detection. DETAILED DESCRIPTION AND PREFERRED EMBODIMENTS THE OVERALL SEPARATION MEANS EXAMPLE 1 In general, first and second dimensional separation means are provided in the current invention to partition a complex mixture of analyte substances. The first dimensional separation means segregates the analyte stream relative to some physio-chemical parameter in order to vary the elution times of said analyte molecules. The second dimensional separation means further segregates the outflowing first dimensional analyte stream into a predetermined number of isoelectric point (pH) ranges. To achieve said first and second dimensional separation means, a series of apparatus are provided and combined in the current invention as described hereto. A means of delivering a mobile phase fluid is provided. Said mobile phase fluid possesses suitable buffering and other physio-chemical characteristics, of the type typically found in biological purification media, and is delivered at a pressure suitable to overcome the pressure drop during flow through a first and second dimension separation apparatus. The corrosive nature of said biological purification media on metal parts is such that a means of delivering the mobile phase without contact to the precision moving parts of a high pressure pumping means is desirable, but optional. FIG.1 illustrates the relationship between the various components of the overall method in one embodiment in which the mobile phase used for the method maintains a non-varying composition with time and isolates said mobile phase from the high pressure pumping means. In a strategy commonly used in the art of high performance biological chromatography, a syringe-like cylindrical reservoir assembly (4) is provided to encapsulate the full volume of mobile phase that will be needed for the complete separation. Said assembly is comprised of a cylindrical reservoir(5) and is divided into two chambers (7a, 7b) by a piston (6). A high pressure pumping means (3) is attached to the upstream chamber (7a) of said reservoir with a high pressure tube (2b) in order to deliver non-corrosive drive fluid under high pressure to said chamber from a drive fluid storage reservoir (1) through tube (2a). Such a non- corrosive drive fluid is typically pure water without buffers or other solutes. Said mobile phase fluid, containing solutes for biological media, is filled into the downstream chamber (7b) of said reservoir for delivery, under pressure, to the first dimension separating means (10) through a sample introduction apparatus (8) described in detail below. Optionally, the high pressure mobile phase pumping means (3) can be a bio- compatible type pump in which the moving parts of the pumping mechanism are designed to withstand the corrosive nature of typical biological buffer mobile phases. In this optional means of mobile phase delivery, the mobile phase reservoir assembly (4) is directly connected to the upstream side of a bio-compatible pumping means (3) and the outflow of said pumping means is directly connected to a sample introduction apparatus (8) described in detail below. A means for the introduction of an analyte sample having an equivalent volume or a smaller volume than the volume of the first dimension stationary phase matrix is provided (8). In one preferred embodiment of the current invention, said means of sample introduction is provided by a sample reservoir or sample loop (9) that can be incorporated into the mobile phase fluid stream, downstream from said mobile phase reservoir by a mechanical means equivalent to the existing art of two position four- way valves, such that the sample reservoir or sample loop in one said position is fluidly connected to an inlet and outlet port for sample loading into said sample reservoir or sample loop. In the same said valve position, the mobile phase stream is fluidly connected on the upstream side to the downstream side of the reservoir or pumping means with tube (2c), and is connected on the downstream side to the first dimension separating means with tube (2d). When said sample introduction means is mechanically or manually repositioned into the opposite valve position, the same said sample loop or sample reservoir is incorporated into the mobile phase stream where the upstream side of said sample loop or reservoir is connected to the mobile phase stream. A means is provided to contain a first dimension partitioning stationary phase matrix in a cylindrical chamber or column (10) of the type typically used for chromatography, so as to allow the passage of the mobile phase over said partitioning matrix in a controlled manner and to retain said matrix within said cylindrical chamber or column while allowing for outflow on the downstream side of said cylindrical chamber. Said cylindrical chamber or column (10) is provided with sufficient strength and fluid integrity so as to resist the level of pressure needed to move the mobile phase through both the first and second dimension separating means, while maintaining the partitioning matrix against pressure compression. In the embodiment of the current invention, in which the first dimension partitioning matrix is a packed microporous or nanoporous granular size exclusion medium of the type typically used for high resolution separation of biological macromolecules, such as proteins, an analyte molecule having a large stokes radius corresponding to a high molecular weight will be excluded from all or most of the fractional volume of the interior of said nanoporous granule by steric hindrance. Analyte molecules with smaller stokes radii will be excluded from a diminishing fraction of the interior volume of said nanoporous granule. The result is that the total internal and external volume that a given analyte molecule can access determines the time required to traverse said column. This is known as the art of size exclusion chromatography. In this preferred embodiment, a single mobile phase reservoir provides a non-varying mobile phase composition. If the complex analyte sample is introduced into the size exclusion means so as to have all molecules in a single small volume relative to the separating volume, then the residence time and elution time for each analyte molecule will be proportional to its stokes radius and consequently its molecular weight, with the largest analyte molecules having the shortest residence and elution times. The downstream outlet of the first dimension separating means(10) is directly connected to the first stage (11) of the second dimension separating means with a high pressure connecting tube (2e), such that diffusion of the analytes is minimized within the connecting tube. FIG. 1 further depicts, in a schematic fashion, the arrangement and functioning of three stages of binary pH range segregation within the second dimension separation means. It is to be understood that, in FIG. 1 , for the purpose of description of the overall functioning of the second dimension separation means, only a single binary pH range segregating manifold assembly (11) is depicted for the first stage of separation. Likewise, only two different binary pH range segregating manifold assemblies (18) and four different binary pH range segregating manifold assemblies (19) are depicted for the second and third stages of separation respectively. It is further to be understood that in all preferred embodiments of the current invention, a plurality of binary pH range segregating manifold assemblies are combined within a single manifold plate and a plurality of manifold plates are combined to form a single separation stage and any number of separation stages can be serially combined to encompass the second dimension separation means. In this respect, identical binary pH range segregating manifold assemblies are fluidly connected in parallel as will be described below. The analyte stream of the first dimension separation means, located within high pressure connecting tube (2e), is delivered to the first stage (11) of the second dimension separation means so as to enter the laminar analyte flow channel on the anodic side (16a), and the laminar analyte flow channel on the cathodic side (16b) of a semi-permeable pH buffering membrane (15). Said anodic side laminar flow channel (16a) is bounded on the anode (12) face by a different type of semi- permeable membrane (14a) that allows the passage of low molecular weight solutes having molecular weights below the molecular weight of any of the analyte molecules to be analyzed, typically about 500-2000 Daltons. Similarly, the cathodic side laminar flow channel (16b) is bounded on the cathode (13) face by another low molecular weight cut-off (about 500-2000 Daltons) semi-permeable membrane (14b). As the flowing analyte stream traverses the laminar analyte flow channels, (FIG.1) a direct current electrical field potential is applied between the anode(12) and the cathode (13) so as to induce electrophoretic migration of the analyte molecules within said channels. The semi-permeable pH buffering membrane (15), having immobilized buffering moieties with a specific aggregate pK value, provides a barrier to said electrophoretic migration (pH 7 for example in the first stage as depicted in FIG.1). Those analyte molecules with isoelectric points below (more acidic than) the aggregate pK value will develop a negative charge within the buffering zone of said membrane (15) and those analyte molecules with isoelectric points above (more basic than) the aggregate pK value will develop a positive charge within the buffering zone of said membrane (15). Analyte molecules that start out in the anodic side laminar flow channel (16a) will move through said semi-permeable membrane into the cathodic side laminar flow channel (16b) only if they retain a net positive charge in the buffering zone of said membrane (15) and will continue to migrate toward the cathode (13). Conversely, analyte molecules that start out in the cathodic side laminar flow channel (16b) will move through said semi-permeable membrane into the anodic side laminar flow channel (16a) only if they retain a net negative charge in the buffering zone of said membrane (15) and will continue to migrate toward the anode (12). Analyte molecules that start out in a laminar space pH range that contains their isoelectric point will develop a net repulsive charge within the buffering zone of the semi-permeable membrane and will be prevented from passing through the membrane. The migrating analyte molecules will either lose their net charge when outside of the buffering zone of said buffering membrane (15) or will be prevented from further migration towards the electrodes by the low molecular weight cut-off membranes (14a) and (14b). The result of the said electrophoretic migration is a segregation of the entire analyte stream into binary pH range fractions during a single passage through a manifold assembly. Additionally, non-amphoteric ionic solutes within the analyte stream will electrophoretically migrate through said low molecular weight cutoff membranes (14a) and (14b) into a space between said low molecular weight cutoff membranes and the electrodes (12) and (13), thus segregating said non- amphoteric ionic solutes from the analyte stream in a process analogous to the art of electrodialysis. The outflowing acidic side laminar flow stream (16a), segregated during passage through the first stage (11) so as to contain all the analyte molecules with isoelectric points below the aggregate pK value of the first stage buffering membrane (pH 7 in the example), is connected to both sides (16e) and (16f) of the second stage (18) binary segregating manifold shown on the left in FIG. 1 that has a pH 4.5 aggregate pK buffering membrane in the example. These analyte streams are further segregated into binary pH range fractions during a single passage through the second stage (18) in the same fashion as in the first stage. The outflowing analyte laminar flow stream (16e) will contain only analyte molecules with an isoelectric point below pH 4.5. The analyte laminar flow stream (16f) will contain only analyte molecules with an isoelectric point between pH 4.5 and pH 7. Likewise, analyte laminar flow stream (16d) will contain only analyte molecules with an isoelectric point between pH 7 and pH 9.5 while analyte laminar flow stream (16c) will contain only analyte molecules with an isoelectric point above pH 9.5. The two binary segregating manifolds shown in the second stage (18) share a common cathode, thus have reversed polarities but function in exactly the same manner. In the third stage of segregation (19) four binary segregating manifolds and four concomitant aggregate pK buffering membranes divide the analyte stream into eight ever narrower pH segregating ranges in laminar flow channels (16g, 16h, 16i, etc). The number of pH ranges and stages of separation providing multi-channel binary pH range segregation is limited only by the accuracy of titration of the immobilized pH buffering moieties. FIG.2 illustrates the relationship between the various components of the overall method in another embodiment; wherein the mobile phase the first dimension solution based separation method uses a time variant (gradient) mobile phase composition and the mobile phase fluid is isolated from the pumping means in the same fashion as is shown in FIG.1. An example of a first dimensional separating means using such a time variant mobile phase composition includes but is not limited to ion exchange chromatography; wherein the mobile phase composition would increase in soluble ionic strength with time, forming a gradient, in order to elute ionically bound analyte molecules from a stationary phase within the first dimensional separating means. In a similar fashion to FIG.1, two syringe-like reservoir assemblies (4, 4") are provided to encapsulate the full volume of mobile phase that will be needed for the complete separation. Typically, the composition of the mobile phase fluid in each said reservoir would vary and represent two extremes of composition that can be combined in a controlled fashion by varying the flow rate of delivery of each said mobile phase fluid. Said reservoir assemblies (4, 4') are each comprised of cylindrical reservoirs (5, 5') divided by pistons (6, 6') into two chambers (7a, 7b) and (7c, 7d) respectively. High pressure pumping means (3,3') withdraw a non-corrosive drive fluid from a drive fluid storage reservoir (1) through tubes (2a, 2f). Said high pressure pumping means are attached to the upstream chambers (7a, 7c) of each cylindrical reservoir, and use high pressure tubes (2b, 2g) to deliver said non-corrosive drive fluid at a high pressure to said upstream chambers. Such a non-corrosive drive fluid is typically pure water without buffers or other solutes. The downstream chambers (7b, 7d) of each cylindrical reservoir are filled with said mobile phase fluids containing solutes for biological media. The mobile phase fluids are delivered, under pressure, to a mobile phase mixing means (20) through high pressure tubes (2h, 2i), wherein the mobile phase fluids from each reservoir are combined in a predetermined time varying ratio. The option exists of using bio-compatible type pumps for each high pressure mobile phase pumping means in the time variant (gradient) embodiment depicted in FIG. 2 in a similar fashion to that described for the non-varying (isocratic) mobile phase composition embodiment. Said optional arrangement would connect each reservoir assembly (4, 4') to the upstream side of bio-compatible versions of each said high pressure pumping means (3, 3') and connect the downstream side of each said pumping means (3, 3") to the aforementioned mobile phase mixing means (20) wherein the mobile phase fluids from each reservoir are combined in a predetermined time varying ratio in a similar fashion to the arrangement depicted in FIG. 2. In either option, said combined mobile phase fluid is delivered through a high pressure tube (2c) to a sample introduction means (8) that is identical to that depicted in FIG. 1, having a sample reservoir or sample loop (9) that can be incorporated into the mobile phase fluid stream in the fashion described in FIG.1. High pressure tube (2d) delivers the mixed mobile phase fluids containing the incorporated sample to the upstream side of the first dimensional separation means (10). The configuration of the first and second dimension separation means depicted in FIG.2 is identical to that described for the arrangement depicted in FIG. 1. In the preferred embodiments of the current invention in which the first dimension partitioning matrix is a packed microporous granular media using ionic, hydrophobic or other physio-chemical differences in the analyte molecules for partitioning, said analyte molecules will be distributed in retention time in the first dimensional separating means in accordance with the partition coefficient of each analyte molecule between the stationary phase matrix and the mobile phase either under non- varying mobile phase composition (isocratic conditions) as in FIG. 1 or under varying mobile phase composition (gradient conditions) FIG. 2. The downstream outlet of the first dimension separating means (10) is directly connected to the first stage (11) of the second dimension separating means with a high pressure connecting tube (2e), such that diffusion of the analytes is minimized within the connecting tube. FIG. 2 further depicts (in a schematic fashion identical to the arrangement and functioning of three stages of binary pH range segregation shown in FIG. 1) a second dimension separation means. It is likewise to be understood that, in FIG. 2, as in FIG. 1 , only a limited number of binary pH segregating manifold assemblies are shown and all preferred embodiments of the current invention use a plurality of binary pH range segregating manifold assemblies combined within a single manifold plate and a plurality of manifold plates combined to form a single separation stage and that any number of separation stages can be serially combined to encompass the second dimension separation means. The description of the arrangement and functioning of the second dimension separation means in FIG. 2 is identical to the above description for FIG. 1 in all respects. FIG.3 illustrates a preferred configuration of fluid channels, electrical connections and overall shape for a single cathode plate assembly (21). Typically said cathode plate assembly would be about 2-20 cm in height and about 2-20 cm in width overall. Each said cathode plate assembly consists of a rigid electrically conductive cathode plate (13) and a rigid non-electrically conductive support plate (22). The thickness of said cathode plate (13) and said support plate (22) are identical and typically would be about 0.05-0.2 cm. In a preferred embodiment of the present invention, said cathode plate (13) is molded from a rigid electrically conductive polymer composite so as to provide a series of channels on each side when assembled with various other components into a manifold assembly. Said molding material should provide for a high modulus of elasticity in bending, a low coefficient of thermal expansion and a conducting surface that is resistant to electrochemical corrosion. It is desirable that such electrode material be easily formed into relatively complex shapes. Such rigid electrically conductive polymeric molding materials include but are not limited to compression molded high performance composite thermoplastics such as polyimide or polyetherimide resins with graphite fiber reinforcement and with or without a corrosion resistant conducting surface coating of stainless steel, platinum or gold. If included, the corrosion resistant conducting surface coating can be limited in area to the plurality of areas in the electrode that correspond in lateral dimension to the channels. In a further preferred embodiment, said rigid support plate (22) is molded from similar resins formulated and reinforced so as to be non-electrically conductive. Said cathode plate (13) is further provided on each side with a plurality of horizontal depressions or grooves (17), typically about 0.05-0.5 cm in height and slightly less in width than the width of said cathode plate. The depth of said horizontal depressions or grooves (17) would typically be about 10% to 30% of the thickness of said cathode plate (13). The horizontal depressions or grooves (17) on opposite faces of said cathode plate are offset in vertical position relative to one another. The amount of said offset is such that the overall plurality of horizontal depressions or grooves on one side of said cathode plate is shifted relative to the horizontal depressions or grooves on the opposite face by an amount equivalent to one half the typical on center vertical spacing of said horizontal depressions or grooves. This offset arrangement of horizontal depressions or grooves, when aligned with the various other components of the overall manifold assembly provides for maximum flexibility in the arrangement of supply and outlet channels for the multiple analyte streams. Said cathode plate (13) provides for a means of electrical connection to other cathode plates within a single stack and a means of supply and return of cooling fluid to each face of said cathode plate. Said means are provided by two extension tabs of the cathode plate on the upper right corner and on the lower left corner beyond the body of the cathode plate with corresponding slots in the non-electrically conductive support plate (22). A transverse hole (29) in the upper right corner tab provides for a supply of cathode cooling fluid to be delivered through a series of vertical depressions or grooves (26) to the plurality of cooling fluid channels formed from said horizontal depressions or grooves (17) upon assembly of the various components into a manifold assembly (11). A transverse hole is defined here as a hole perpendicular to the face of a plate that penetrates from on side to the other. Such transverse holes can align along an axis so as to form transverse channels. Likewise, a transverse hole (27) in the lower left corner tab provides for the return of cathode cooling fluid to be collected from the same said series of cooling fluid channels connected through additional vertical depressions or grooves (26). The aforementioned means of electrical connection for said cathode plate (13) is provided by an additional transverse hole (31) in the upper right corner extension tab above transverse hole (29). The transverse hole (31) aligns with other transverse holes on adjacent cathodes to provide for the insertion of a conducting rod, such that a plurality of cathodes are connected to each other and to a source of electrical potential upon assembly of a complete stack of manifolds into a single stage of binary separation (see FIG. 9). The rigid non-electrically conductive support plate (22) fits tightly around said cathode plate (13) so as to support and electrically isolate said cathode plate from other components of the apparatus. Transverse holes (28), (30) and (32) in said support plate (22) align, on assembly, with matching transverse holes in anode plate (12) as shown in FIG. 4, thus providing equivalent cooling fluid supply, return and electrical connections for said anode plate (12). The transverse holes (28), (30) and (32) in said support plate (22) ensure electrical and fluid isolation of said anode plate (12) from said cathode plate (13). FIG. 3 further illustrates a plurality of transverse holes (23) in a vertical column on the left end of support plate (22). Each hole (23) is aligned vertically with the upper half of each horizontal depression or groove (17) on the front face of cathode plate (13) and the lower half of each horizontal depression (17) on the back face of cathode plate (13) (see FIG. 3B). Said holes (23), on assembly with the various other components of the apparatus, provide uninterrupted transverse channels through all or some portion of a complete stack of manifolds in a single stage of binary separation. Said transverse channels, formed in part by holes (23), provide a means of integrating and combining the outflowing analyte streams from all equivalent cathodic side laminar flow channels (16b) as shown in FIG. 1 from all previous stage binary segregating manifold assemblies. Said integrating transverse channels formed on assembly, are referred to as "cathodic analyte stream collection and distribution manifolds" and represent, in all cases, the more basic pH range analyte stream from each given segregating manifold. Transverse holes located in various other components of the apparatus that are contiguous with transverse holes (23) and provide for transverse channels which will also be labeled (23) in this description. Likewise, the plurality of transverse holes (24) in a vertical column, located to the right of and above each concomitant transverse hole (23), are vertically aligned with the lower half of each horizontal depression or groove (17) on the front face of cathode plate (13) and the upper half of each horizontal depression (17) on the back face of cathode plate (13) (see FIG. 3B). Said holes (24) form a related set of transverse integrating channels on assembly with various other components of the apparatus. Said formed channels are referred to as "anodic analyte stream collection and distribution manifolds" and by contrast collect in all cases the more acidic pH range analyte stream from each given segregating manifold. FIG. 3 also illustrates a plurality of transverse holes (25) in a vertical column on the right end of support plate (22). Said plurality of transverse holes (25) provide fluid channels connecting the equivalent outflowing cathodic analyte streams on both sides of said cathode plate assembly (21) (see FIG. 3C) for subsequent connection to the "cathodic analyte stream collection and distribution manifolds" of the downstream binary pH range separation stage. In one embodiment of the current invention, cathode support plate (22) also provides a means of connection of equivalent fluid channels in some adjacent columns or rows of transverse channels as is required to integrate equivalent outflowing pH range analyte streams. Said means of connection is provided by non- penetrating short vertical or horizontal hole to hole depressions or grooves in said cathode support plate (22) similar to horizontal depressions or grooves (17) or vertical depressions or grooves (26). Said hole to hole connecting depressions or grooves are formed in said cathode support plate (22) only as required in occasional positions and thus are not depicted in FIG. 3. FIG. 3B illustrates a vertical cross sectional view of cathode plate (13). This diagram depicts how transverse channel (23) for collecting the "cathodic (more basic) analyte stream" and transverse channel (24) for collecting the "anodic (more acidic) analyte stream" are aligned in the vertical direction with grooves (17) in the lower position on the front side of cathode plate (13) and with grooves (17) in the upper position on the back side of cathode plate (13). FIG. 3C illustrates a cut away isometric cross sectional view of cathode plate (13), buffering membrane support plate (36) and anode plate (12) and their respective support plates showing how they assemble to provide collection channels for the cathodic analyte stream that route said streams around the end joints of the cathode support plate (22) where it butts against the cathode support plate (22) of the downstream stage. This downstream cathode support plate is shown by a dotted line. Spacer gaskets (40) (see FIG. 6A) and (47) (see FIG. 7B) are also depicted. This diagram depicts how the outflowing cathodic (more basic) analyte stream in the channel aligned with front side groove (17) in the cathode plate (13) passes through short transverse channel (25) and connects to the outflowing cathodic analyte stream in slot (48) of spacer gasket (47). The said combined cathodic analyte streams continue through aligned transverse hole (39) in the buffering membrane support plate (36) and aligned transverse hole (42) in spacer gasket (40) into a short connecting groove (65) in the face of anode support plate (24). The said combined analyte stream continues through said short connecting groove (65) to empty into the "cathodic analyte stream collection and distribution manifold" formed on assembly (as shown) by the various aligned transverse holes (23) in buffering membrane support plate (36), spacer gasket (40), anode support plate (24) as well as other layers not shown. It should be noted that all "cathodic analyte stream collection and distribution manifolds" are contiguous through all layers having equivalent binary separation aggregate pK values and thus pool all equivalent cathodic analyte streams. This is also true for all "anodic analyte stream collection and distribution manifolds" In a manner similar to FIG. 3, FIG. 4 illustrates a preferred configuration of fluid channels, electrical connections and overall shape for a single anode plate assembly (33). Anode plate assembly (33) is of identical size, shape and composition as cathode plate assembly (21) shown in FIG. 3. Anode plate assembly (33) consists of anode plate (12) and anode support plate (34). In a similar manner to transverse hole (31) on cathode plate (13), a transverse hole (32) located on an extension tab on the upper left side of anode plate (12) provides a means of electrical connection, on assembly, between all said anode plates within a single stage of binary pH range segregation and electrical connection to a source of electrical potential. In an analogous manner to FIG. 3, transverse hole (28) located on an extension tab on the lower right corner of anode plate (12) provides for a supply of anode cooling fluid to be delivered through a series of vertical depressions or grooves(35) to the plurality of cooling fluid channels formed from the horizontal depressions or grooves (17). Likewise, transverse hole (30) located on the upper left corner below hole (32) provides an outlet channel for anode cooling fluid to be collected from said plurality of cooling fluid channels formed from the horizontal depressions or grooves (17). Non-conducting anode support plate (34) fits tightly around said anode plate (12) so as to support and electrically isolate said anode plate (12) from other components of the apparatus. Transverse holes (27) and (29) in the lower left and upper right corners of anode support plate (34) align with transverse holes (27) and (29) in cathode plate (12) so as to provide, on assembly, cathode cooling fluid supply and outlet channels. FIG. 4 further illustrates a plurality of transverse holes (23) and (24) on the right side of anode support plate in vertical rows that align with similarly named transverse holes (23) and (24) in cathode support plate (22). Said transverse holes (23) and (24) form, on assembly, the aforementioned outflowing analyte stream transverse integrating channels referred to as "cathodic analyte stream collection and distribution manifolds" and "anodic analyte stream collection and distribution manifolds" In a similar fashion to cathode support plate (22), in one embodiment of the current invention, anode support plate (34) also provides a means of connection of equivalent fluid channels in some adjacent columns or rows of transverse channels as is required to integrate equivalent outflowing pH range analyte streams. Said means of connection is likewise provided by non-penetrating short vertical or horizontal hole to hole depressions or grooves in said anode support plate (34) similar to horizontal depressions or grooves (17) or vertical depressions or grooves (35). Said hole to hole connecting depressions or grooves are formed in said anode support plate (34) only as required in occasional positions and thus, as in FIG. 3, are not depicted in FIG. 4. FIG. 5 illustrates a single semi-permeable buffering membrane assembly (15) comprised of a molded elastomeric buffering membrane support plate(36), a plurality of semi-permeable buffering membranes (37) and a macroporous reinforcement screen (38). Said reinforcement screen (38) consists of an inert open weave fabric screen or net with at least 90% open cross-sectional area that is embedded into said molded support plate (36) during molding to provide dimensional stability without substantially reducing the cross-sectional area of the openings provided for the buffering membranes. Typically, said molded support plate (36) would be about 2-20 cm in height and about 2-20 cm in width overall and is provided with a plurality of horizontal transverse slots and a plurality of transverse holes that correspond in size and position with the matching plurality of horizontal depressions or grooves (17) and plurality of transverse holes (23) and (24) located on cathode plate (13) and anode plate (12). The thickness of said molded support plate (36) typically would be about 0.05-0.2 cm. In a preferred embodiment of the present invention, said support plate (36) is molded from elastomeric or semi-elastomeric polymers to include but not limited to the classes; Polysiloxane, Polyisoprene, Polyisobutylene and Polysulfide. Said elastomeric polymers or semi-elastomeric polymers have physical properties that provide for the easy and tight sealing of molded support plate (36) against various other components of the apparatus. Additionally, molded support plate (36) provides for fluid connection channels across the thickness of said molded support plate using a plurality of transverse holes (39) located inboard on the right and left side of molded support plate (36). Said transverse holes (39) are, in some instances, aligned with similar transverse holes, such as the plurality of transverse holes (36), in various other components of the apparatus. Said transverse holes (39) provide fluid channels that distribute analyte streams only from one side of said molded support plate (36) or cathode support plate (22) to another within a single segregating manifold assembly. Said fluid channels so formed by the plurality of transverse holes (39) do not provide a means of integrating multiple equivalent analyte streams as do transverse holes (23) and (24). FIG. 5 further illustrates a plurality of semi-permeable buffering membranes (37) that are cast into the pre-formed plurality of horizontal slots in support plate(36). Each said semi-permeable buffering membrane (37) is cast into an aforementioned slot so as to have the exact thickness of support plate (36), to have a smooth surface flush with said support plate (36) and to have macroporous reinforcement screen (38) embedded within said semi-permeable buffering membrane (37) so as to provide reinforcement such that the dimensional stability of the porous matrix is maintained without substantially reducing the porosity or capacity of the matrix to provide an immobilized buffering means. Each said semi-permeable membrane (37) consists of a porous matrix having multiple buffering chemical moieties immobilized in specific titrated ratios within said matrix such that a particular aggregate pK value and buffering capacity is established in each said membrane (37). In a preferred embodiment of the present invention, said buffering chemical moieties are of the general class of acrylamide derivatives known as acrylamido-buffers and are co-polymerized into said semi-permeable membrane along with acrylamide and bis-acrylamide or other acrylamido derivatives of acrylamide during polymerization. Said semi-permeable membrane (37) establishes the primary means of binary pH range segregation of the analyte streams. Each aggregate pK formulation of a specific semi-permeable membrane (37) comprises a different pre-determined composition of acrylamido-buffers combined with acrylamide, bis-acrylamide and or acrylamido derivatives of acrylamide. It is a desired feature of the present invention that equivalent semi-permeable membranes (37) having the exact same aggregate pK value be cast from the same formulation batch of monomeric precursors at the same time in order to maintain uniformity in the subsequent semi-permeable membranes (37) and resultant pH range analyte fractions. FIG. 6A illustrates a spacer gasket (40) to be located between the back side of molded support plate (36) and low molecular weight membrane (14a) that is adjacent to the front side of anode plate assembly (33) This assembly is shown in FIG. 8. Said spacer gasket (40) is comprised of a thin, semi-rigid sheet of non-conducting polymeric material of uniform thickness, wherein said thickness varies by not more than 5 % over the entire surface of said spacer gasket. An example of such a material includes but is not limited to polyethylene terephthalate sheet. Typically, said spacer gasket (40) would be about 2-20 cm in height and about 2-20 cm in width overall. Said spacer gasket (40) is also provided with a plurality of horizontal transverse slots (41) and a plurality of transverse holes (23) and (24) that correspond in size and position with the matching plurality of horizontal slots (containing semi-permeable membranes (37)) and the plurality of transverse holes (23) and (24) in molded support plate (36) (see FIG. 5), anode support plate (34) (see FIG. 4) and cathode support plate (22). The thickness of said molded support plate (36) typically would be about 0.01-0.1 cm. Spacer gasket (40) provides a means of forming and maintaining the laminar flow channels, (16a) for example, on an anodic side of semi-permeable membrane (37) as illustrated schematically in FIG. 1. Said horizontal transverse slots (41) are generally aligned with said semi-permeable membranes (37) so as to bring the analyte stream located in said transverse slots (41) in very close contact with the entire surface of semi-permeable membrane (37), anode plate (12) and cathode plate (13) thereby optimizing electrophoretic migration of amphoteric analyte molecules across said semi-permeable membrane (37). Each individual horizontal transverse slot (41) provides a means of connecting one "cathodic analyte stream collection and distribution manifold" from a previous stage to said horizontal transverse slot (41) by a narrow extension of said slot so as to fluidly connect each said horizontal transverse slot (41) with one of the plurality of transverse holes (39) in the upstream side of molded support plate (36) (FIG. 5), thus providing inflowing cathodic analyte streams to said horizontal transverse slot (41). Likewise, a narrow extension of said slot (41) on the downstream side (right side in FIG. 6A) provides a means of connecting the acidic pH segregated outflowing analyte stream in said slot (41) to a subsequent stage of segregation by fluidly connecting each said horizontal transverse slot (41) with one "anodic analyte stream collection and distribution manifold" located on the downstream side of the manifold assembly through a horizontal groove in anode support plate (34) thus establishing a continuous flowing path for said analyte stream on the anodic side of semi-permeable membrane (37). Spacer gasket (40) further provides a transverse hole (42) that is not fluidly connected directly or indirectly to transverse slot (41) and said anodic side laminar flow channels. Said transverse hole (42) aligns with some transverse holes (39) to provide a means of connecting certain unrelated cathodic side laminar flow channels to the "cathodic analyte stream collection and distribution manifolds" In a similar fashion FIG. 6B illustrates a spacer gasket (43) be located between the back side of molded support plate (36) and the low molecular weight membrane (14b) that is adjacent to the front side of cathode plate assembly (21). Said spacer gasket (43) is likewise about 2-20 cm wide and about 2-20 cm high and comprised of a thin, semi-rigid sheet of non-conducting polymeric material of uniform thickness, wherein said thickness [does not vary by greater] varies by no more than 5 % over the entire surface of said spacer gasket. Spacer gasket (43) with slots (44) provides a means of forming and maintaining the laminar flow channels, (16b) for example, on a cathodic side of semi-permeable membrane (37) as illustrated schematically in FIG. 1. A plurality of transverse holes (23) and (24), located on the left end of spacer gasket (43), correspond in size and position with the "cathodic analyte stream collection and distribution manifolds" and the "anodic analyte stream collection and distribution manifolds" respectively. Spacer gasket (43) provides a means for the inflow of an anodic analyte stream and an outflow of the basic pH segregated outflowing analyte stream in said slot (44) to a "cathodic analyte stream collection and distribution manifold" in the downstream side of the separation manifold in the same manner as described in FIG. 6A for spacer gasket (40). FIG. 7A illustrates a spacer gasket (45) with a plurality of transverse slots (46) to be located between the front side of molded support plate (36) as shown in FIG.5 and the back side of anode plate assembly (33) as shown in FIG. 4. Spacer gasket(45), of the same dimensions as spacer gasket (40) (FIG. 6A), provides a means for the inflow of an anodic analyte stream and an outflow of the acidic pH segregated analyte stream in each said slot (46) to an "anodic analyte stream collection and distribution manifold" in the downstream side of the separation manifold in the same manner as described in FIG. 6A for spacer gasket (40). Finally, FIG. 7B illustrates a spacer gasket (47) with a plurality of transverse slots (48) to be located between the front side of molded support plate (36) as shown in FIG. 5 and the back side of cathode plate assembly (21) as shown in FIG. 3. Spacer gasket (47), of the same dimensions as spacer gasket (40) (FIG. 6A), provides a means for the inflow of a cathodic analyte stream and an outflow of the basic pH segregated analyte stream in each said slot (48) to a "cathodic analyte stream collection and distribution manifold" in the downstream side of the separation manifold in the same manner as described in FIG. 6A for spacer gasket (40). EXAMPLE 2 ONE BINARY SEGREGATING MANIFOLD ASSEMBLY FIG. 8 illustrates the relationship between the various components of one binary segregating manifold assembly (11) in an exploded view. In this illustration a binary segregating manifold assembly with the cathode on the front side and the anode on the back side is shown. The binary segregating manifolds on each side of the one shown will have a reverse polarity because they share electrodes with this binary segregating manifold. The arrangement of components in these reversed polarity assemblies is a mirror of what is depicted in FIG. 8. A single binary segregating manifold assembly such as (11) is comprised of a cathode plate assembly (21) at the front of the overall exploded view depicted in FIG. 8. Said cathode plate assembly (21) is illustrated in detail in FIG. 3. This is followed (in order from front to rear in FIG. 8) by a low molecular weight cutoff semi-permeable membrane (14b), and spacer gasket (47) shown in detail in FIG. 7B. Said spacer gasket (47) is positioned with respect to the cathode plate assembly (21) such that slots (48) (FIG. 7B) are aligned with the corresponding grooves (17) in cathode plate (13) (FIG. 3), which is a part of said cathode plate assembly (21). Next, a semi-permeable buffering membrane assembly (15), illustrated in detail in FIG. 5 is positioned adjacent to the cathode side spacer gasket (47) and similarly aligned such that the individual buffering membranes (37) correspond to the slots (48) in said spacer gasket (47). A spacer gasket (40) shown in detail in FIG. 6A is positioned against said semi-permeable buffering membrane assembly (15) with slots (48) (FIG. 6A) in spacer gasket (40) aligned with said individual buffering membranes (37) in said semi-permeable buffering membrane assembly (15). A second low molecular weight cutoff semi-permeable membrane (14a) is then positioned between said anode side spacer gasket (40) and the anode plate assembly (33) shown in detail in FIG. 4. The horizontal depressions or grooves (17) in anode plate assembly (33) (FIG. 4) are likewise aligned with said slots (48) in anode side spacer gasket (40). Upon assembly of a binary segregating manifold assembly such as (11), illustrated in FIG. 8, the cathode plate assembly (21) and the anode plate assembly (33) provide for transverse fluid channels (27), (28), (29), and (30) (that have been previously described in FIG.3 and FIG. 4). Said fluid channels pass through the entire binary segregating manifold assembly (11). Said transverse fluid channels are further aligned with similar transverse fluid channels in other binary segregating manifold assemblies stacked within a single stage as shown in FIG. 9. The purpose of the transverse fluid channels (27), (28), (29), and (30) is to provide a path of delivery and removal of electrode cooling fluid to the cathode plate (13) and the anode plate (12). Additionally, the binary segregating manifold assembly (11), illustrated in FIG. 8, provides a transverse hole (31) passing completely through said manifold assembly for the purpose of connecting the cathode plate (13) using an electrical conductor rod (51) shown in FIG.9 with other cathode plates within a stacked manifold plate assembly and to a source of negative electrical potential. Likewise, a transverse hole (32) passing completely through said manifold assembly (11) provides an electrical connecting means for anode plate (12) using an electrical conductor rod (50), also shown in FIG.9, connected to other anode plates within a stacked manifold plate assembly and to a source of positive electrical potential. Each binary segregating manifold assembly (11), is provided with a plurality of semi-permeable membranes, located in semi-permeable buffering membrane assembly (15) described in detail in FIG.5, that on assembly with said spacer gasket (47) and spacer gasket (40), divide said manifold assembly into two sets of laminar flow channels such as (16a) and (16b), for instance, as shown in FIG.1. Said laminar flow channels run parallel to the direction of the continuously flowing mobile phase. The inflowing mobile phase enters both sets of laminar analyte flow channels simultaneously on each side of said semi-permeable buffering membrane through the series of transverse supply holes previously described in FIGS.3-7B. Conversely, the outflowing pH segregated mobile phases exit separately from each laminar flow channel of said semi-permeable buffering membrane through a series of transverse outlet holes also previously described in FIGS. 3-7B. All said transverse supply and outlet holes connect to a limited number of other lateral or transverse supply and collection channels such as "cathodic analyte stream collection and distribution manifolds" and "anodic analyte stream collection and distribution manifolds" both located between stages in a pattern that is designed to provide for the distribution of the various analyte stream pH range fractions to the required zones of equivalent binary segregating manifold assemblies in subsequent separation stages as described in general in FIG. 1. Additionally, the low molecular weight cutoff semi-permeable membranes (14a) and (14b), shown in FIG. 8, provide for the containment of analyte molecules (having molecular weights above the cutoff of about 500-2000 Daltons) within the laminar analyte flow channels while allowing the free flow of small charged and uncharged molecules below the cutoff of about 500-2000 Daltons through said low molecular weight cutoff semi-permeable membranes (14a) and (14b) into the spaces that contain the electrodes. Said low-molecular weight cutoff semi-permeable membranes (14a) and (14b) typically have a minimal thickness as required to provide for the semi- permeable containment. Examples of such a material include but are not limited to the type of cellulose membrane known as low molecular weight cutoff benzoylated cellulose dialysis tubing. It is a desired feature of the current invention that equilibration of the analyte stream by isoelectric point segregation occurs entirely within the residence time of said analyte stream in a given pH segregation stage. Said segregation can be achieved by a process of rap/of equilibration in order to maintain overall high flow rates through all stages of separation. In one preferred embodiment, the physical dimensions of the current invention are such that rapid equilibration is achieved by the combination of; (1 ) a large surface area for the semi-permeable buffering membrane relative to the volume of the laminar flow channels, (2) a short transfer distance and (3) a maximum potential electrical field gradient provided over (4) a small electrode spacing. FIG.9 illustrates a further assembly of individual manifold assemblies into a plurality of separation stages. Two such separation stages (49) and (49") are shown. It is to be understood that the number of separation stages, such as (49) and (49'), that can be combined end to end, as depicted in FIG.9, is theoretically unlimited. Although there are no intrinsic limits to the number of stages that can be used, depending on the technology, practical or physical limitations may serve to limit the number of stages. In the preferred embodiment for rapid equilibration, individual manifold assemblies, such as (11), shown in FIG.8 that are located within a single separation stage such as (49) or (49'), as illustrated in FIG.9, are further assembled into a compressed stack. Said individual manifold assemblies, in the current embodiment are depicted as (11) in FIG. 9 and are assembled as described in FIG.8 except that every other individual manifold assembly (11) is reversed in polarity and shares an electrode with the adjacent individual manifold assembly. Thus, an individual manifold assembly (11) can be construed to run from the vertical centerline of one anode plate to the vertical centerline of the next cathode plate. The adjacent individual manifold assembly then runs from the same said cathode plate centerline to the next anode plate centerline. This arrangement is also depicted schematically in FIG.1. The result of this arrangement is that the anodic side laminar flow analyte streams, shown as (16a), (16d), (16e) and (16g) in FIG.1 , and the cathodic side laminar flow analyte streams shown as (16b), (16c), (16f), (16h) and (16i) in FIG.1, are located on different sides of the semi-permeable buffering membrane assembly (15) in adjacent individual manifold assemblies. The direction of analyte flow from the first dimensional segregation means (10), or from some previous separation stage of the overall second dimension separation means, is shown for separation stage (49) as arrow (52) and as arrow (52') for the downstream separation stage (49') in FIG.9. Alignment of the various connecting fluid channels between separation stages is provided by overlap between the various layered components of individual manifold assemblies in one separation stage and the layered components of manifold assemblies in other separation stages. For example, the transverse holes (24) located in the downstream side of spacer gasket (40) FIG. 6A, spacer gasket (45) FIG. 7A and anode support plate (34) FIG. 4 in separation stage (49), overlap to form connecting channels with similar transverse holes (24) in the upstream side of spacer gasket (43) FIG. 6B, spacer gasket (47) FIG. 7B, and cathode support plate (22) FIG. 3 within the adjacent individual manifold assembly in the downstream separation stage (49'). ' Additional overlapping, in this fashion of the layered components of manifold assemblies in the two separation stages transforms said plurality of aligned transverse holes (24) into an uninterrupted "anodic analyte stream collection and distribution manifold" over the entire zone of the separation stage having equivalent semi-permeable buffering membrane assemblies (15). FIG.9 further illustrates for each separation stage (49) and (49'), two electrical conductor rods (50), (51) and (50'), (51') respectively. Said electrical conductor rods (50) and (50') run through all of the transverse holes (32) (FIG.4) in all the anode plates a single separation stage such as (49) or (49'). Said electrical conductor rods (50) and (50') provide a means of electrical of connection of all said anode plates in a single separation stage to a source of positive electrical potential. Likewise, said electrical conductor rods (51) and (51") run through all of the transverse holes (31) (FIG.3) in all the cathode plates a single separation stage such as (49) or (49"). Said electrical conductor rods (51) and (51') provide a means of electrical of connection of all said cathode plates in a single separation stage to a source of negative electrical potential. FIG.9 further illustrates for each separation stage, (49) for instance, two transverse channels (28) and (29), that run through all individual manifold assemblies in said single separation stage. Said transverse channels (28) and (29) provide a means of supply and delivery of cooling fluid to each anode plate and cathode plate respectively in said separation stage. In order to prevent a short circuit path for electrical potential between the anodes and cathodes, said transverse channels (28) and (29) are physically and electrically isolated from one another. Likewise, FIG.9 illustrates for each separation stage, (49) for instance, two transverse channels (27) and (30), that run through all individual manifold assemblies in said single separation stage. Said transverse channels (27) and (30) provide a means of return and collection of cooling fluid from each cathode plate and anode plate respectively in said separation stage. Again, in order to prevent a short circuit path for electrical potential between the anodes and cathodes, said transverse channels (27) and (30) are physically and electrically isolated from one another. Said anode and cathode cooling fluid streams also provide for the removal of low molecular weight (about 500-2000 Daltons) charged and uncharged molecules that pass through the semi-permeable low molecular weight membranes (14a) and (14b). This design provides for exceptionally high electrical fields by eliminating the resultant high current and concomitant coulombic heating caused by small molecule electrolytes in the analyte stream that can reduce the effective electrical field strength. Exceptionally high electrical fields minimize the equilibration time. EXAMPLE 3 OVERALL CONFIGURATION OF BINARY SEGREGATING CHANNELS IN STAGES FIG. 10 illustrates in an exploded view, an overall configuration of the second dimension separation means. Said overall second dimension separation means is comprised of a plurality of separation stages (49, 49', 49"and 49'") corresponding to separation stages n=0, n=1 , n=2 and n = n respectively. Also, said overall second dimension separation means provides for an upstream pressure cap (53), and a combined downstream pressure cap and outlet nozzle array plate (54), and a means (not shown) of enclosing all said separation stages between said pressure caps (53) and (54) so as to maintain said separation stages at a pressure or pressures above atmospheric pressure as required to move said analyte stream through said overall second dimension separation means. In addition, said overall second dimension separation means, shown in FIG. (10), is comprised of a plurality of movable fraction collection plates (55) for collecting simultaneous fractions from each final outlet nozzle and, finally, a plurality of solid phase extraction array assemblies (56) for immobilizing some part of the outlet stream analyte molecules for detection. It is to be understood that only four separation stages (49, 49', 49"and 49'") of many are shown in FIG. 10. It is to be further understood that any number of separation stages can be combined to form the overall second dimension separation means. The overall analyte stream, from the first dimension separation means (10), enters said overall second dimension separation means through a port in upstream pressure cap (53) and is distributed across the entire upstream face of separation stage n=0 (49). Said overall analyte stream has a general direction of flow from each said upstream separation stage into subsequent downstream separation stages. Said general direction of flow of the analyte stream from stage to stage is depicted by arrows (52, 52" and 52'}. The combination of a plurality of binary pH segregating channels distributed vertically within a single manifold assembly and a plurality of said manifold assemblies stacked horizontally to form a binary separation stage, as shown in FIG. (9), provides for an array of channels in two dimensions as viewed from the inlet or outlet faces of said binary separation stage. As the pH range of each binary analyte stream narrows by stages, the number of analyte species (isoelectric point values) within each pH range is reduced in a binary fashion. Assuming that the capacity of a single channel remains constant, a preferred configuration of said channels is such thata// channels in the initial segregating stage provide for a single initial buffering membrane aggregate pK value, pH 7 for instance as shown in FIG. (1). As illustrated in FIG. 10 and Table 2 below, in the case wherein there are 32 channels in each manifold and 32 manifolds that are stacked to form a single stage, 1024 parallel channels in separation stage n=0 (49) (with 1024 equivalent semi- permeable buffering membrane channels (37)) form a single zone (A) in said separation stage n=0 (49). Said 1024 channels would segregate the total analyte stream into two pH ranges, greater than pH7 and less than pH 7 in the example shown in FIG. (1). The basic analyte streams (greater than pH 7 in this example) would be collected and combined in the array of transverse channels formed by aligned transverse holes (23) and vertical channels (not shown) in the downstream periphery of separation stage n=0 (49) to form an uninterrupted "cathodic analyte stream collection and distribution manifold" over the entirety of zone (A). Similarly, the acidic analyte streams (less than pH 7 in this example) would be collected and combined in an identical array of transverse and vertical channels formed by aligned transverse holes (24) and vertical channels (not shown) in the downstream periphery of said separation stage n=0 (49) to form an uninterrupted "anodic analyte stream collection and distribution manifold" over the entirety of zone A. Said anodic and cathodic distribution manifolds are unconnected. Viewed from the outlet side, initial separation stage n=0 (49) consists of a single zone (A) of similar binary pK value buffering membranes providing two pH range analyte streams in unconnected distribution manifolds extending over the entire zone. The second separation stage n=1 (49') consists of two types of binary pH segregating membranes (37) (at pH 4.5 and 9.5 in this example) dividing the analyte stream into four different pH ranges. Said pH 4.5 segregating membranes are located in zone (B) comprised of all 32 vertical channels on the leftmost 16 manifold assemblies of the stack in said separation stage n=1 (49"). Likewise, the pH 9.5 segregating membranes are located in zone (C) on the rightmost 16 manifold assemblies of said separation stage n=1 (49"), thus dividing the 32 x 32 array into two zones, 16 channels wide by 32 channels high. Said anodic distribution manifolds between separation stage n=0 (49) and separation stage n=1 (49') distribute the integrated acidic analyte stream (less than pH 7 in this example) collected from separation stage n=0 (49) into only those channels that comprise zone (B) of separation stage n=1 (49'). Likewise, said cathodic distribution manifolds between separation stage n=0 (49) and separation stage n=1 (49') distribute the integrated basic analyte stream (greater than pH 7 in this example) collected from separation stage n=0 (49) into only those channels that comprise zone (C) of separation stage n=1 (49'). As in separation stage n=0 (49), a similar array of transverse channels and vertical channels in the downstream periphery of separation stage n=1 (49') form uninterrupted "cathodic and anodic analyte stream collection and distribution manifolds". In this case however, transverse holes (23 and 24) in only the particular anode support plate (34) that is located between zone (B) and zone(C) are not formed, thus providing a barrier that divides said "cathodic and anodic analyte stream collection and distribution manifolds" into separate zones corresponding to zone (B) and zone(C) and limiting the distribution of said outflowing analyte streams from separation stage n=1 (49') to said zones (B) and zone(C) on the upstream face of separation stage n=2 (49"). In the third separation stage n=2 (49") (with binary pH segregating membranes (37) at aggregate pK values of pH 3.25, pH 5.75, pH 8.25 and pH 10.75 for example) each 16 wide by 32 high zone (B) and zone(C) from separation stage n=1 (49") would be divided into two zones each 16 channels wide by 16 channels high. The pH 3.25 segregating membranes located in the uppermost and leftmost zone(D) of the stack, the pH 5.75 located in the lowermost and leftmost zone (F) and so forth. Again downstream side transverse holes (23 and 24) in the particular anode support plate (34) located between zone (B) and zone(C) are not formed. In addition vertical connections between the uppermost 16 channels of each manifold assembly and the 16 lowermost channels of each manifold are not formed, thus dividing the connected "cathodic and anodic analyte stream collection and distribution manifolds" into 4 zones corresponding to zones (D-G) 0 Subsequent stages subdivide the 32 x 32 array into zones, each containing fewer and fewer equivalent binary pH segregating membranes (37) in a similar fashion (i.e. 8 wide by 16 high, 8 wide by 8 high, 4 wide by 8 high, etc.) Likewise, the arrays of interconnected collecting channels in the periphery of each stage have limiting 5 barriers between corresponding zones on the upstream face of the subsequent separation stage, thus limiting distribution of analyte streams to single downstream zones. In the given example, a final separation stage wherein n = 10 (49'"), a single channel of each species of binary pH segregating membrane (37) is provided, forming 2 = 1024 different 1 channel wide x 1 channel high zones, dividing the final 0 outflowing analyte streams into 2 (n+ ' = 2048 final stage pH ranges. TABLE 2 illustrates the number and size of zones within a 32 by 32 array of individual channels (in the current example of 2048 channels per stage) for each stage of separation. 5 Stage # equivalent pK # equivalent pK # equivalent pK value Zone array Zone array n value manifolds value channels zones (channels (channels /stage /manifold /stage wide) high) 0 32 32 1 (A) 32 32 1 16 32 2 (B,C) 16 32 2 16 16 4 (D,E,F,G) 16 16 3 8 16 8 8 16 Stage # equivalent pK # equivalent pK # equivalent pK value Zone array Zone array n value manifolds value channels zones (channels (channels /stage /manifold /stage wide) high) 4 8 8 16 8 8 5 4 8 32 4 8 6 4 4 64 4 4 7 2 4 128 2 4 8 2 2 256 2 2 9 1 2 512 1 2 10 1 1 1024 (2 n ) 1 1 EXAMPLE 4 5 A MEANS OF COLLECTING ANALYTE FRACTIONS A means is illustrated in FIG. 10 of one preferred embodiment of collecting fractions following said final separation stage n = 10 (49'") of multi-channel segregation. In the given example, wherein said final separation stage n = 10 (49'") of 0 multi-channel segregation consists of 1024 channels and divides the analyte stream into 2048 final stage pH ranges, there are two outlets for each binary pH segregating membrane (37) species. This results in an array of outlet channels 64 wide by 32 high. 5 The aforementioned downstream pressure cap and outlet nozzle array plate (54) provides an array of 64 vertical columns (about 0.6-6mm on center) of outlet nozzles (57) and 32 horizontal rows (about 1.2-12mm on center) of outlet nozzles(57), i.e. the spacing between the rows of nozzles, typically is twice that between columns of nozzles. Said outlet nozzle array plate (54) is additionally provided with 32 horizontal 0 surface grooves (59) located between each row of horizontal outlet nozzles (57), running the full width of said end cap and extending to each edge of said end cap. Said horizontal surface grooves (59) are spaced equally between each said row of horizontal outlet nozzles (57) to provide an open channel on the face of outlet nozzle array plate (54) to the atmosphere. Detail A of FIG. 10 illustrates a close up view of the outlet nozzle array plate (54), a single movable fraction collection plate (55) and a single solid phase extraction array (56). The aforementioned plurality of movable fraction collection plates (55) have a width and height corresponding to the width and height of the outlet nozzle array in outlet nozzle array plate (54). Said movable fraction collection plates (55) are provided with an array of transverse channels or micro-capillary tubes (58) corresponding in number and position with the array of nozzles on said outlet nozzle array plate (54). Said channels or micro-capillary tubes (58) being normal to the surface of said movable fraction collection plates (55) and passing completely through said movable fraction collection plates (55) are thus parallel and aligned with said outlet nozzles (57) in outlet nozzle array plate (54). Detail A of FIG. 10 further illustrates a close up view showing said transverse channels or micro-capillary tubes (58) in said movable fraction collection plate (55). The diameter of each said transverse channel or micro-capillary tube (58) is comparable but not necessarily identical to the diameter of said outlet nozzle (57). Each said transverse channel or micro-capillary tube (58) may be a separate removable micro-capillary tube held in place within said movable fraction collection plate (55) or a transverse channel permanently molded into said movable fraction collection plate (55). The thickness of said movable fraction collection plate (55) is variable as required to provide transverse channels or micro-capillary tubes (58) of variable internal volume. Typically said movable fraction collection plate (55) is about 0.2-2.0 cm in thickness. A means is provided of rapidly transporting and positioning said movable fraction collection plate (55) directly against the outlet nozzle array plate (54) such that each of the 2048 outlet nozzles (57) in the example, is aligned with one of the 2048 transverse channels or micro-capillary tubes (58) in said movable fraction collection plate (55). A further means is provided to bring said movable fraction collection plate (55) into close contact with said outlet nozzle array plate (54). Said close contact forms a tight seal between each transverse channel or micro-capillary tube (58) and each outlet nozzle (57) such that the outflowing analyte stream from each said outlet nozzle (57) collects in a corresponding transverse channel or micro-capillary tube (58), thus collecting a single fraction of the outflowing stream from each of the outlet nozzles (2048 in the example) simultaneously. The time of contact between said movable fraction collection plate (55) and outlet nozzle array plate (54) determines the fraction collection time of the analyte streams and therefore the fraction volume in each said transverse channel or micro-capillary tube (58). Said time of contact can be varied from less than the time required for filling the transverse channels or micro-capillary tubes (58) to more than the time required for filling said transverse channels or micro-capillary tubes (58). In the case wherein said time of contact is greater than the time required for filling said transverse channels or micro-capillary tubes (58), a portion of the analyte streams in each said transverse channel or micro-capillary tube (58) will pass completely through the said movable fraction collection plate (55). At the end of the said fraction collection time of an array of single fractions from each said outlet nozzle (57), a means is provided to rapidly shift the position of said movable fraction collection plate (55) vertically relative to said outlet nozzle array plate (54) along the face of said array plate (54) and without losing contact with said array plate (54). Said rapid vertical shift unseals said transverse channels or micro-capillary tubes (58) in movable fraction collection plate (55) from said corresponding outlet nozzles (57) in outlet nozzle array plate (54). Said rapid vertical shift further aligns the transverse channels or micro-capillary tubes (58) with said horizontal surface grooves (59) located between rows of nozzles on said outlet nozzle array plate (54). Said horizontal surface grooves (59) provide a means venting said transverse channels or micro-capillary tubes (58) to the atmosphere at the point of contact of each said transverse channel or micro-capillary tube (58) and each said outlet nozzle (57), thus isolating that portion of the analyte stream within each said transverse channel or micro-capillary tube (58). A further means is provided to rapidly move said movable fraction collection plate (55) in a horizontal direction parallel to said horizontal surface grooves (59) as shown by arrow (63) in FIG. 10, thus maintaining venting for all said channels or tubes in said fraction collection plate until said fraction collection plate is completely clear of said outlet nozzle array plate (54). This arrangement prevents the loss or cross contamination of analyte fractions between channels or tubes. Simultaneously with said horizontal movement (63) of the analyte fraction filled movable fraction collection plate (55), a second, empty movable fraction collection plate (55) is positioned in it's place, such that the transverse channel or micro- capillary tubes (58) in said empty plate (55) are initially aligned with said horizontal surface grooves (59). Once the empty movable fraction collection plate (55) is in place horizontally, it is rapidly shifted vertically so as to align the transverse channels or micro-capillary tubes (58) in said empty movable fraction collection plate (55) with outlet nozzles (57) on said outlet nozzle array plate (54) to form a tight seal, between each transverse channel or micro-capillary tube (58) and each outlet nozzle (57) thus filling the empty fraction collection plate in the current example, with an array of 2048 second fractions from each analyte stream. A plurality of filled and empty movable fraction collection plates (55) are provided with a means of stacking and de-stacking said movable fraction collection plates (55) in order to place said plurality of movable fraction collection plates (55) in position for fraction collection and storage over the full time required for a complete sample analysis. EXAMPLE 5 MEANS OF DETECTING ANALYTES A means is provided to simultaneously detect analytes in the outflowing analyte stream from each outlet nozzle (57) of said outlet nozzle array plate (54) located at the downstream face of the final separation stage n = 10 (49'") of the overall second dimension multi-channel apparatus as shown in FIG. 10. Said simultaneous means of detection can be continuous at the time of analyte stream outflow from the second dimension separation means or said means of detection can be delayed in time or in rate of detection thus differing from the rate of outflow of said analyte stream. In a preferred embodiment of the current invention, such a means of detection must achieve several requirements; (1 ) said means of detection must detect analytes in all channels essentially at the same time or serially within a short time, (2) said means of detection must detect analytes non-destructively or with a minimum of sample in order to provide, for further analysis, the aforementioned fractions in said channels or micro-capillary tubes (58) and (3) a means of detection must detect all analytes equally and with a high degree of sensitivity. As illustrated in FIG. 10, including Detail A, the aforementioned plurality of solid phase extraction array assemblies (56) provide a means of immobilizing some portion of the outlet stream analyte molecules for detection. Each said solid phase extraction array assembly (56) consists of a rigid support plate (60) with a plurality of transverse channels (61) normal to the surface of said rigid support plate (60), forming a horizontal and vertical array (64 columns by 32 rows in the current example). Solid phase extraction array assembly (56) also consists of a membrane (62) bonded to a face of said rigid support plate (60) such that said membrane (62) closes off one end of each said transverse channel (61). Said membrane (62) provides a means of extracting and immobilizing analyte molecules simultaneously from each said analyte stream as said analyte streams pass through said transverse channels (61). In the embodiment of the invention wherein the analyte molecules being segregated are amphoteric molecules and in particular said analyte molecules are protein or peptide molecules, said membrane (62) is comprised in whole or part of one of several membrane materials known to bind protein or peptide molecules such as polyvinylidene difluoride and nitrocellulose. The width and height of said solid phase extraction array assembly (56) is about 2-20cm. The thickness of said rigid support plate (60) of solid phase extraction array assembly (56) is about 0.01 -0.1 cm. In addition, the thickness of said membrane (62) of solid phase extraction array assembly (56) is about 0.001 -0.01 cm. It is a desired feature of the current invention that said membrane (62) of said solid phase extraction array assembly (56) be so formed as to have a minimal and exceptionally uniform thickness while still providing immobilizing capacity sufficient to immobilize the entirety of said analyte molecules in each said analyte stream over the time interval selected for said analyte molecule immobilization. Said desired feature provides for concentrating and placing said extracted and immobilized analyte molecules into a single plane within the area of said membrane (62) that covers said transverse channel (61), thus enhancing subsequent detection by those methods that utilize the art of time of flight mass spectroscopy. Said transverse channels (61) correspond in position and alignment with said channels or micro-capillary tubes (58) in said movable fraction collection plate (55). A means is provided of vertically transporting and positioning said solid phase extraction array assembly (56) directly against the downstream face of movable fraction collection plate (55) such that each of the 2048 transverse channels (61) for instance, in the example shown in FIG. 10, is aligned with one of the 2048 transverse channels or micro-capillary tubes (58) in said movable fraction collection plate (55). Once aligned, said solid phase extraction array assembly (56) is tightly sealed against said movable fraction collection plate (55) so as to allow the analyte stream in each channel or micro-capillary tube (58) in movable fraction collection plate (55) to flow into and through each corresponding transverse channel (61) impinging and immobilizing said analyte molecules through chemical adsorption onto the area of said membrane (62) that covers said transverse channel (61). At the end of the selected sampling and immobilization period for the analyte streams through transverse channels (61) onto membranes (62), a means is provided to rapidly shift the position of the fully adsorbed solid phase extraction array assembly (56) vertically relative to said movable fraction collection plate (55) and replace said fully adsorbed solid phase extraction array assembly (56) with a new solid phase extraction array assembly (56) as shown in FIG. 10 by arrow (64). The selected sampling and immobilization period for the analyte streams in said solid phase extraction array assemblies (56) can be chosen to be less than, equal to, or greater than, the selected period of fraction collection in said movable fraction collection plate (55) as desired. This provision allows maximum flexibility in allocating analyte streams between the detection means and the fraction collection means as required. A plurality of fully adsorbed and new solid phase extraction array assemblies (56) are provided with a means of stacking and de-stacking said solid phase extraction array assemblies (56) in order to place a sufficient number of said solid phase extraction array assemblies (56) in position for sample extraction and immobilization during the full time required for a complete first and second dimension sample analysis. The same said means of stacking and de-stacking said solid phase extraction array assemblies (56) can be used as a means of positioning said fully adsorbed solid phase extraction array assembly (56) in position, as required for the chosen method of subsequent detection. Said immobilized analytes can then be analyzed by several methods. A preferred method of detection and analysis of macromolecular analytes such as proteins extracted onto a solid phase matrix, is matrix assisted laser desorption ionization mass spectroscopy, in particular infrared laser desorption mediated time of flight mass spectroscopy (IR-MALDI-TOF). Said infrared laser desorption mediated time of flight mass spectroscopy (IR-MALDI-TOF) has been described in the prior art. In the same said preferred method of detection and analysis of macromolecular analytes, the aforementioned means of placing said fully adsorbed solid phase extraction array assembly (56) into position for infrared laser desorption mediated time of flight mass spectroscopy (IR-MALDI-TOF), can further provide a means for placing said fully adsorbed solid phase extraction array assembly (56) in contact with a means of cooling said fully adsorbed solid phase extraction array assembly (56). The purpose of said means of cooling said fully adsorbed solid phase extraction array assembly (56) is to freeze the residual analyte mobile phase, or otherwise absorbed solutions, in membrane 62) that covers each said transverse channel (61). Said freezing of said absorbed solutions in membrane 62) occurs before entry of said fully adsorbed solid phase extraction array assembly (56) into the required vacuum chamber means of the infrared laser desorption mediated time of flight mass spectroscopy (IR-MALDI-TOF) apparatus. Said freezing provides a required means of embedding said analyte molecules into a energy absorbing matrix as required for infrared laser desorption. There are several advantages to the aforementioned preferred method of analyte detection using IR-MALDI-TOF. Laser desorption methods in general require a very small quantity of analyte embedded in a solid phase energy absorbing matrix. In this preferred method of analyte detection, the energy-absorbing matrix can be water ice, ethanol ice, glycerol or other media that are readily compatible with a protein analyte, thereby greatly simplifying the subsequent analysis. Said prior art of laser desorption time of flight mass spectroscopy provides for the pulsed desorption of matrix and analyte in a very short time frame (between 200 picoseconds and several nanoseconds) followed by an electrostatic acceleration of the ionized analyte molecules through a vacuum drift tube into a detector with typical total flight times for macromolecules in the range of 10-1000 microseconds. Also, a single pulse can be focused to an area of the target matrix of about 200 microns in diameter and can be subsequently redirected onto other target locations at high speed. These characteristics provide for the rapid scanning and analysis of a solid phase extracted analyte stream immobilized onto an array grid such as the solid phase extraction array assembly (56), with a cycle time in the example given, of less than one second up to several seconds for all 2048 positions in the grid. Said cycle time is well within the time frame of significant sample composition variation in a typical analyte stream and thus represents a sufficient sampling time for detection of analyte peaks. The output signal for time of flight mass spectroscopy provides a measure of the mass (molecular weight) of the intact analyte molecule with a resolution typically of one part in 300 to 500 (full width half maximum) and a measure of the relative intensity of the mass peak corresponding to the amount of analyte. Analyte peaks of a few femtograms can be detected and the outline of potentially overlapping or hidden analyte "spots" can be elucidated by the appearance of additional peaks in the mass spectrogram, when scanning an area of a two dimensional map. While the precision of measurement of mass resolution using time of flight mass spectroscopy is relatively low compared with sector or resonant methods of mass spectroscopy, it is considerably better than the approximate 5% error in molecular weight typically found in 2D gel electrophoresis. This level of mass resolution can greatly help in the unambiguous identification of individual analytes even when only partially separated. Additional means of detection include all the established methods of immobilized sample detection to include but not limited to; optical density detection, dynamic and static fluorescence detection, conductivity detection, electrochemical detection, dynamic and static light scattering detection and other forms of mass spectroscopic detection. In a separate embodiment of the current invention in order to provide said additional means of detection, the solid phase extraction array assemblies (56) are replaced with a second movable fraction collection plate (55) so as to collect the analytes over a period of time in a compact array of corresponding analyte stream sampling volumes. In this embodiment of the current invention, analyte stream sampling volumes collected and stored in a plurality of transverse channels (61) within second movable fraction collection plate (55), can be analyzed in real time or delayed time using a means of moving and positioning said second movable fraction collection plate (55) into a position for transferring said sampling volumes into a detector micro- flow cell or an electrospray ionization means for the introduction of analyte molecules into one of several alternate forms of mass spectroscopy. In order to provide said separate embodiment using a second movable fraction collection plate (55) a means of scanning or multiplexing said detector or electrospray ionization means to the various transverse channels (61) in a serial fashion, Said alternate forms of mass spectroscopy include but are not limited to, the prior arts of tandem quadrapole mass spectroscopy and Fourier transform ion cyclotron resonance mass spectroscopy.

Claims: The invention claimed is: 1. An apparatus for the solution based separation and detection of amphoteric substances in complex mixtures in a plurality of dimensions in a continuously flowing fluid segregating component analytes to a predetermined level of resolution in multiple channels comprising: a high pressure pumping mechanism to provide a flow of fluid containing amphoteric substances; a high pressure tube connected at one end to said high pressure pumping mechanism; a mobile phase reservoir assembly attached at an opposing end of said high pressure tubing; a second length of high pressure tubing attached to downstream end of said mobile phase reservoir; a sample loop incorporated into said second length of high pressure tubing to introduce an analyte sample having a volume at most equivalent to a volume of a first dimension stationary phase matrix; a first dimension separation means connected to an opposing end of said second length of high pressure tubing; a third length of high pressure tubing connected to a downstream outlet of said first dimension stationary phase matrix; a first stage binary segregating manifold assembly of a second dimension separating means attached to an opposing end of said third length of high pressure tubing; a plurality of intermediate stages of said second dimension separating means connected to said first stage; a final separation stage connected to last intermediate stage of said plurality of intermediate stages; a collection stage fluidly connected to said final separation stage; and a detection stage adjacent to a downstream face of said collection stage. 2. The collection stage according to claim 1 , further comprising: a collection mechanism having capabilities to collect outflowing discrete fractions of a plurality of binary pH range isoelectric point segregated analyte streams following said final separation stage for subsequent analysis and use. 3. The detection stage according to claim 1 , further comprising: a detection mechanism for amphoteric substances in a plurality of binary pH range isoelectric point segregated analyte streams providing real time detection of outflowing analytes from said final separation stage. 4. The first dimension separation means according to claim 1 , selected from the group consisting essentially of a free flowing mobile phase and a stationary (immobile) phase that is in contact with all parts of the mobile phase providing a means of selective partition of analytes between the mobile phase and the stationary phase to include strong and weak cation exchange chromatography, strong and weak anion exchange chromatography, size exclusion chromatography, hydrophobic interaction chromatography, hydrophilic interaction chromatography, hydroxyapatite chromatography, capillary electrophoresis, dye interaction chromatography, fast performance liquid chromatography, reverse phase chromatography, perfusion chromatography and non specific affinity chromatography. 5. The first dimension separation means according to claim 1 , further comprising: a column of sufficient strength and fluid integrity to resist levels of pressure required to move mobile phase through a first dimension stationary phase matrix and said second dimension separating means; and a first dimension stationary phase matrix of a microporous or a nanoporous granular size exclusion medium in an interior of said column for high resolution separation of biological macromolecules having a large stokes radius corresponding to a high molecular weight are excluded from a fractional volume of said interior of said nanoporous granule by steric hindrance. 6. The first stage binary segregating manifold assembly of said second dimension separating means according to claim 1 , further comprising: a cathode plate assembly; a low molecular weight cutoff semi-permeable membrane positioned adjacently to a rear of said cathode plate assembly; a cathode side spacer gasket positioned adjacently to a rear of said low weight cutoff semi-permeable membrane having a position with respect to said cathode plate assembly so that each slot is aligned with a corresponding groove in said cathode plate assembly; a semi-permeable buffering membrane assembly positioned adjacently to the rear of said cathode side spacer gasket having an alignment such that each individual buffering membrane corresponds to a slot in said cathode side spacer gasket; an anode side spacer gasket is positioned against a rear side of said semi-permeable buffering membrane assembly with slots of said anode side spacer gasket aligned with individual buffering membranes in said semi-permeable buffering membrane; a second low molecular weight cutoff semi-permeable membrane adjacently to the rear of said anode side spacer gasket; and an anode plate assembly having multiple depressions or grooves aligned with a multiple of slots in said anode side spacer gasket positioned adjacently to the rear of said second low molecular weight cutoff semi-permeable membrane. 7. The cathode plate assembly according to claim 6, further comprising: a portion of rigid non-electrically conductive support plate to support said cathode plate while electrically isolate said cathode plate from other components of the apparatus; a plurality of transverse holes in a vertical column on a left end of said portion of rigid non-electrically conductive support plate aligned vertically with a lower half of said horizontal depressions providing cathodic analyte stream collection and distribution manifolds through at least some portion of a complete stack of manifolds integrating and combining outflowing analyte streams from all equivalent cathodic side laminar flow channels collecting a more basic pH range analyte stream; a second plurality of transverse holes in a vertical column to the right of and above each concomitant transverse hole where said second plurality of transverse holes are vertically aligned with an upper half of each horizontal depression forming a related set of anodic analyte stream collection and distribution manifolds on the assembly with various other components of said apparatus and collect the more acidic pH range analyte stream from each given segregating manifold; a third plurality of transverse holes in a vertical column on a right end of said portion of rigid non-electrically conductive support plate support plate provide fluid channels connecting an equivalent outflowing cathodic analyte streams on both sides of said cathode assembly for subsequent connection to said cathodic analyte stream collection and distribution manifolds; a fourth plurality of transverse holes in an upper left corner of said portion of rigid non- electrically conductive support plate having a top hole providing a means of electrical connection to other anode plates within a single stack by alignment with other transverse holes on adjacent anode plate assemblies to allow insertion of a conducting rod that acts as source of electric potential while also connecting said anode plate assemblies to one another while a bottom hole returns anode cooling fluid; and a fifth transverse hole diagonally positioned from said fourth plurality of transverse holes in a lower right portion of said portion of rigid non-electrically conductive support plate supplying anode cooling fluid. 8. The cathode assembly plate of claim 7, further comprising: a portion of rigid electrically conductive cathode plate surrounded by said portion of rigid non-electrically conductive support plate having a height of about 2 to 20 centimeters, a width of about 2 to 20 centimeters with a thickness of about 0.05 to 0.2 centimeters; a plurality of horizontal depressions or grooves on opposite faces of said upper portion of rigid electrically conductive cathode plate offset in vertical position relative to one another such that said plurality of horizontal depressions on one side of said cathode plate is shifted relative to said horizontal depressions on an opposite face by an amount equivalent to one half of typical on center vertical spacing of said horizontal depressions having a height of about 0.05 to 0.5 centimeters with a width less than said width of said upper portion of rigid electrically conductive cathode plate with a depth of about 10 percent to 30 percent of said thickness of said upper portion of rigid electrically conductive cathode plate; a set of tabs extending beyond the body of said cathode plate upwards from an upper right corner of said cathode plate above said horizontal depressions with another extending downwards from a lower left corner of said cathode plate below said horizontal depressions; a plurality of transverse holes where said tab in said upper right corner has at least an upper transverse hole and a lower transverse hole with said upper transverse hole providing a means of electrical connection to other cathode plates within a single stack by alignment with other transverse holes on adjacent cathode plate assemblies to allow insertion of a conducting rod that acts as source of electric potential while also connecting said cathode plate assemblies to one another while said lower transverse hole supplies cathode cooling fluid through a series of vertical depressions to a plurality of cooling fluid channels formed from said horizontal depressions; and a third transverse hole in said set of tabs diagonally positioned from said plurality of transverse holes in said upper right corner providing for a return of cathode cooling fluid to be collected from said series of cooling fluid channels connected through additional vertical depressions. 9. The anode plate assembly according to claim 6, further comprising: a portion of rigid non-electrically conductive support plate to support said anode plate while electrically isolate said anode plate from other components of the apparatus; a plurality of transverse holes in a vertical column on a right end of said portion of rigid non-electrically conductive support plate aligned vertically with a lower half of said horizontal depressions providing cathodic analyte stream collection and distribution manifolds through at least some portion of a complete stack of manifolds integrating and combining outflowing analyte streams from all equivalent cathodic side laminar flow channels collecting a more basic pH range analyte stream; a second plurality of transverse holes in a vertical column to the right of and above each concomitant transverse hole where said second plurality of transverse holes are vertically aligned with an upper half of each horizontal depression forming a related set of anodic analyte stream collection and distribution manifolds on the assembly with various other components of said apparatus and collect the more acidic pH range analyte stream from each given segregating manifold; a third plurality of transverse holes in a vertical column on a left end of said portion of rigid non-electrically conductive support plate support plate provide fluid channels connecting an equivalent outflowing anodic analyte streams on both sides of said anode assembly for subsequent connection to said anodic analyte stream collection and distribution manifolds; a fourth plurality of transverse holes in an upper right corner of said portion of rigid non-electrically conductive support plate having a top hole providing a means of electrical connection to other cathode plates within a single stack by alignment with other transverse holes on adjacent cathode plate assemblies to allow insertion of a conducting rod that acts as source of electric potential while also connecting said cathode plate assemblies to one another while a bottom hole provides for cathode cooling fluid; and a fifth transverse hole diagonally positioned from said fourth plurality of transverse holes in a lower left portion of said portion of rigid non-electrically conductive support plate for returning cathode cooling fluid. 10. The anode assembly plate of claim 9, further comprising: a portion of rigid electrically conductive anode plate surrounded by said portion of rigid non-electrically conductive support plate having a height of about 2 to 20 centimeters, a width of about 2 to 20 centimeters with a thickness of about 0.05 to 0.2 centimeters; a plurality of horizontal depressions or grooves on opposite faces of said upper portion of rigid electrically conductive anode plate offset in vertical position relative to one another such that said plurality of horizontal depressions on one side of said anode plate is shifted relative to said horizontal depressions on an opposite face by an amount equivalent to one half of typical on center vertical spacing of said horizontal depressions having a height of about 0.05 to 0.5 centimeters with a width less than said width of said upper portion of rigid electrically conductive anode plate with a depth of about 10 percent to 30 percent of said thickness of said upper portion of rigid electrically conductive anode plate; a set of tabs extending beyond the body of said anode plate upwards from an upper left corner of said anode plate above said horizontal depressions with another extending downwards from a lower right corner of said anode plate below said horizontal depressions; a plurality of transverse holes where said tab in said upper left corner has at least an upper transverse hole and a lower transverse hole with said upper transverse hole providing a means of electrical connection to other anode plates within a single stack by alignment with other transverse holes on adjacent anode plate assemblies to allow insertion of a conducting rod that acts as source of electric potential while also connecting said anode plate assemblies to one another while said lower transverse hole provides an outlet channel for anode cooling fluid to be collected from said plurality of cooling fluid channels formed from said horizontal depressions; and a third transverse hole in said set of tabs diagonally positioned from said plurality of transverse holes in said lower right corner providing for a supply of anode cooling fluid to be delivered through said series of cooling fluid channels connected through additional vertical depressions. 11. The low molecular weight cutoff semi-permeable membrane and second low molecular weight cutoff semi-permeable membrane of claim 6, further comprising: a membrane allowing for the passage of low molecular weight solutes having molecular weights below the molecular weight of any of a group of analyte molecules to be analyzed in the range of about 500 to 2000 Daltons. 12. The cathode side spacer gasket according to claim 6, further comprising: a semi-rigid sheet of non-conducting polymeric material of a uniform thickness wherein said uniform thickness varies at most by about 5 percent across said cathode side spacer gasket to maintain laminar analyte flow channels. 13. The cathode side spacer gasket according to claim 12, further comprising: a plurality of horizontal transverse slots to form and maintain laminar analyte flow channels wherein said channels are in a lower of two positions as aligned with channels formed into said cathode; a narrow extension protruding from a lower half of a left end of said horizontal transverse slots; and a plurality of transverse holes aligned having an inner column and an outer column positioned at a left side of said cathode side spacer gasket adjacent to said narrow extension. 14. The cathode side spacer gasket according to claim 13, wherein said plurality of horizontal transverse slots form and maintain laminar analyte flow channels having said channels in a higher of two positions as aligned with channels formed into said cathode. 15. The anode side spacer gasket according to claim 6, further comprising: a semi-rigid sheet of non-conducting polymeric material of a uniform thickness wherein said uniform thickness varies at most by about 5 percent across said anode side spacer gasket to maintain a plurality of laminar analyte flow channels. 16. The anode side spacer gasket according to claim 15, further comprising: a plurality of horizontal transverse slots to form and maintain laminar analyte flow channels wherein said channels are in a lower of two positions as aligned with channels formed into said anode; a narrow extension protruding from a lower half of a right end of said horizontal transverse slots; and a plurality of transverse holes aligned having an inner column and an outer column positioned at a right side of said anode side spacer gasket adjacent to said narrow extension. 17. The anode side spacer gasket according to claim 16, further comprising: a third vertical column of transverse holes aligned with a portion of a second plurality of transverse holes on an upstream side of a molded support plate providing a means of collecting certain unrelated cathodic side laminar flow channels to cathodic analyte stream collection and distribution manifold. 18. The anode side spacer gasket according to claim 17, wherein said plurality of horizontal transverse slots form and maintain laminar analyte flow channels having said channels in a higher of two positions as aligned with channels formed into said anode. 19. The semi-permeable buffering membrane assembly of claim 6, further comprising: a molded elastomeric buffering membrane support plate having a height of about 2 to 20 centimeters and a width of about 2 to 20 centimeters with a thickness of about 0.05 to 0.2 centimeters; a macroporous reinforcement screen having an inert open weave fabric screen with at least 90 percent open cross sectional area embedded into said molded elastomeric buffering membrane support plate providing dimensional stability; and a plurality of semi-permeable buffering membranes having uniform composition of a porous matrix with multiple buffering chemical moieties immobilized in specific titrated ratios within said matrix establishing a particular aggregate pK value and buffering capacity flushly cast into a plurality of slots of said molded elastomeric buffering membrane support plate to have a smooth surface flush with said molded elastomeric buffering membrane support plate covering said macroporous reinforcement screen. 20. The molded elastomeric buffering membrane support plate according to claim 19, further comprising: a first plurality of transverse holes in multiple vertical columns on a left side of said molded elastomeric buffering membrane support plate corresponding in size and position with a matching plurality of holes on said cathode plate and said anode plate; a second plurality of holes in multiple vertical columns positioned on a right side of said molded elastomeric buffering membrane support plate opposite from said first plurality of transverse holes; and a plurality of transverse slots horizontally positioned between said first and second pluralities of transverse holes corresponding in size and position to matching horizontal depressions on said cathode plate and said anode plate at a right adjacent position to said plurality of transverse holes. 21. A method for solution based separation and detection of amphoteric substances in a plurality of dimensions in a continuously flowing fluid comprising the steps of: pumping under high levels of pressure a non-corrosive drive fluid from a drive fluid storage chamber to a mobile phase reservoir assembly to deliver the mobile phase to a first dimension separation means through a length of high pressure tubing; introducing an analyte sample, having at most a volume equivalent to a first dimension stationary phase matrix volume, to a second length of high pressure tubing downstream from said mobile phase reservoir by using a sample loop; separating biological macromolecules utilizing a partitioning stationary phase matrix packed with microporous or nanoporous granular size exclusion medium in a column connected to a downstream end of said second length of high pressure tubing; delivering a partitioned outflowing first dimension mobile phase to a first stage binary pH range isoelectric point segregating means through a third length of high pressure tubing connected to a downstream end of said column; and delivering multiple outflowing binary pH range isoelectric point segregated analyte streams into a plurality of further stages of binary pH range isoelectric point segregating means connected to said first stage binary pH range isoelectric point segregating means, ending at a final stage binary pH range isoelectric point segregating means. 22. The method according to claim 1 , further comprising the steps of: collecting a plurality of outflowing binary pH range isoelectric point segregated analyte streams in discrete fractions at an outflow of said final stage binary pH range isoelectric point segregating means using movable fraction collection plates collecting simultaneous fractions from each final outlet nozzle and a plurality of solid phase extraction array assemblies for immobilizing some portion of outlet stream analyte molecules for detection. 23. The method according to claim 1 , further comprising the steps of : detecting analytes using infrared laser desorption mediated time of flight mass spectroscopy in an outflowing analyte stream simultaneously from each final outlet nozzle located at a downstream face of said final stage binary pH range isoelectric point segregating means, where simultaneous of detection can be continuous at the time of analyte stream outflow from said final stage binary pH range isoelectric point segregating means or can be delayed in time or rate of detection to differ from the rate of outflow of said outflowing analyte stream.

Inventors: CHAMPAGNE JAMES T, CHAMPAGNE, JAMES, T.
Applicants: CHAMPAGNE JAMES T, CHAMPAGNE, JAMES, T.
Application year: 2000
Main classification: B01D57
IPC codes: B01D57, G01N27, G01N30