Protein separation by electrophoresis

Abstract

The present method provides methods and apparatus for separating proteins using a series of electrophoretic methods that utilize controlled fractionation and labeling techniques to resolve mixtures of proteins. The samples for each electrophoretic method other than the initial method, contain only a subset of proteins resolved in the preceding method. The methods can be used in a variety of different applications including, creating proteomic databases, comparative expression studies, diagnostics, structure activity relationships and metabolic engineering investigations.

1 - 61 . (Cancelled)  
     
     
         62 . A method for separating a plurality of proteins, comprising performing a capillary isoelectric focusing electrophoresis (CIEF) or capillary zone electrophoresis (CZE) method with a sample containing the plurality of proteins, wherein the CIEF method or CZE method is conducted under conditions such that electroosmotic flow (EOF) is less than or equal to 0.5×10-6 cm 2 /V-s.  
     
     
         63 . The method of  claim 62 , wherein the method comprises performing a CIEF method under conditions such that electroosmotic flow (EOF) is less than or equal to 0.5×10 −6  cm 2 /V-s.  
     
     
         64 . The method of  claim 62 , wherein the method comprises performing a CZE method under conditions such that electroosmotic flow (EOF) is less than or equal to 0.5×10 −6  cm 2 /V-s.  
     
     
         65 . The method of  claim 62 , further comprising detecting at least one protein separated by the CIEF or CZE method.  
     
     
         66 . The method of  claim 65 , further comprising analyzing the at least one protein to determine a chemical or physical characteristic of the at least one protein.  
     
     
         67 . The method of  claim 66 , wherein the chemical or physical characteristic is selected from the group consisting of molecular weight, complete or partial amino acid sequence, isoelectric point, relative or absolute abundance and combinations of the foregoing.  
     
     
         68 . The method of  claim 66 , wherein analyzing is conducted by mass spectrometry.