EXPLOITATION OF A MONOCLONAL-ANTIBODY FOR WEAK AFFINITY-BASED SEPARATION IN CAPILLARY GEL-ELECTROPHORESIS

Weak biospecific recognition has been established for affinity separat ion in high performance liquid chromatography (HPLC). The use of weak affinity chromatography (WAC) has been limited previously by the insuf ficient separation efficiency achieved, allowing only some 1000 plates /m to be obtained. However, it has been shown that chiral drug separat ion can be performed with capillary affinity gel electrophoresis (CAGE ) at considerably improved efficiency as compared with traditional chr omatographic procedures. Our present study demonstrates the potential of weak affinity monoclonal antibodies as a generic method for immunol ogically based separations in capillary electrophoresis. Monoclonal an tibodies were polymerized within a silica capillary and were used for the separation of structurally similar carbohydrate antigens. The resu lts indicate that weak biospecific interactions can be utilized in a C AGE format to produce highly selective separation of the alpha- and be ta-forms of p-nitrophenyl-labeled maltose. It remains to be seen, howe ver, how efficient weak affinity separation in CAGE can be compared wi th affinity HPLC protocols. Details of typical separations and of the preparation of the antibody gel are presented.