QUANTITATION OF SERUM PROTEINS USING 2-D ELECTROPHORESIS

TWO-DIMENSIONAL (2-D) ELECTROPHORESIS IS THE MOST EFFECTIVE METHOD KNOWN FOR RESOLVING COMPLEX MIXTURES OF PROTEINS. AT PRESENT, HOWEVER, THE METHOD IS SELDOM USED FOR QUANTITATIONOF PROTEINS IN HUMAN SERUM BECAUSE OF THREE PROBLEMS: (1) SERUM ALBUMIN INTERFERES WITH THE RESOLUTION OF MOST OTHER SERUM PROTEINS; (2) LOSSES DUE TO THE METHOD ARE NOT EASILY MONITORED; AND (3) COMPUTER SOFTWARE FOR ROUTINE CLINICAL ANALYSES USING 2-D ELECTROPHORESIS IS NOT AVAILABLE. THE RESEARCH PROPOSED IN THIS APPLICATION IS DIRECTED TO- WARD SOLUTION OF THESE PROBLEMS. A METHOD FOR REMOVAL OF ALBUMIN FROM HUMAN SERUM USING AN IMMUNOAFFINITY RESIN WILLDEVELOPED; LOSSES DUE TO THE METHOD WILL BE MONITORED BY BY INCLUSION OF 14C RADIOLABELED STANDARDS, AND SOFTWARE FOR COMPILATION OF CLINICAL DATA WILL BE DEVELOPED FOR THE IBM AT MICROCOMPUTER. EVENTUALLY IT IS HOPED THAT ABOUT 40 SERUM PROTEINS CAN BE MEASURED, TABULATED, AND COMPARED TO REFERENCE VALUES IN A SINGLE 2-D GEL ASSAY TAKING 1 TO 2 WEEKS. IF SO, THIS NEWLY DEVELOPED METHODOLOGY WILL HAVE WIDESPREAD APPLICATION IN DISEASE DIAGNOSIS AND MONITORING OF THERAPEUTIC EFFECTIVENESS.