title: Exploitation of a monoclonal antibody for weak affinity-based separation in capillary gel electrophoresis.
abstract: Weak biospecific recognition has been established for affinity separation in high performance liquid chromatography (HPLC). The use of weak affinity chromatography (WAC) has been limited previously by the insufficient separation efficiency achieved, allowing only some 1000 plates/m to be obtained. However, it has been shown that chiral drug separation can be performed with capillary affinity gel electrophoresis (CAGE) at considerably improved efficiency as compared with traditional chromatographic procedures. Our present study demonstrates the potential of weak affinity monoclonal antibodies as a generic method for immunologically based separations in capillary electrophoresis. Monoclonal antibodies were polymerized within a silica capillary and were used for the separation of structurally similar carbohydrate antigens. The results indicate that weak biospecific interactions can be utilized in a CAGE format to produce highly selective separation of the alpha- and beta-forms of p-nitrophenyl-labeled maltose. It remains to be seen, however, how efficient weak affinity separation in CAGE can be compared with affinity HPLC protocols. Details of typical separations and of the preparation of the antibody gel are presented.
authors.affiliation: Technical Analytical Chemistry, Center for Chemistry and Chemical Engineering, Lund University, Sweden.
authors.forename: H
authors.initials: H
authors.lastname: Ljungberg
authors.forename: S
authors.initials: S
authors.lastname: Ohlson
authors.forename: S
authors.initials: S
authors.lastname: Nilsson
mesh_terms: D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000941:Antigens / Q000302:isolation & purification; D002846:Chromatography, Affinity; D019075:Electrophoresis, Capillary / Q000379:methods*; D008320:Maltose / Q000276:immunology / Q000302:isolation & purification; D009578:Nitrobenzenes / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D009578:Nitrobenzenes; D008320:Maltose