Affinity capillary electrophoresis for the assessment of complex formation between viruses and monoclonal antibodies.
The formation of complexes of human rhinovirus (serotype HRV2 and HRV14) with nonaggregating neutralizing monoclonal antibodies was investigated by affinity capillary electrophoresis. The method is based on preincubation of virus with antibody, followed by CE analysis. At low antibody-to-virus ratios, peaks corresponding to the complexes were broad, pointing to the presence of a heterogeneous population of virions with various numbers of antibodies bound; at a high molar ratio between virus and antibody, the peak became narrow again, indicating saturation of the 60 equivalent viral epitopes with the antibodies being attached bivalently. As SDS was used as an additive in the background electrolyte to allow for separation, its influence on complex formation was investigated. Once formed, HRV2-antibody complexes were found to be stable in the presence of the detergent but complex formation in buffer containing SDS was severely impaired. HRV14-antibody complexes were rapidly dissociated by SDS. The method proved to be useful for a rapid assessment of complex formation and might allow for an estimation of the binding stoichiometry.
Okun
V M
VM
Institute of Analytical Chemistry, University of Vienna, Austria.
Ronacher
B
B
Blaas
D
D
Kenndler
E
E
D000911:Antibodies, Monoclonal / Q000276:immunology*; D019075:Electrophoresis, Capillary; D006801:Humans; D012229:Rhinovirus / Q000276:immunology*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D012967:Sodium Dodecyl Sulfate