Comparison of three methods for competitive binding of monoclonal antibodies The localization of antigenic sites for monoclonal antibodies on Panulirus interruptus hemocyanin

• Published in: Journal of immunological methods Vol. 190; no. 1; pp. 117 - 125
• Main Authors: Perton, Frank G., Dijkema, Jan H., Smilda, Tim, van Uffelen, Bart Erik, Beintema, Jaap J.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 28.03.1996; Elsevier
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibody Specificity; Antigenic region; Binding Sites, Antibody; Binding, Competitive; Biological and medical sciences; Chromatography, Gel; Competitive binding; Crustacea - immunology; Electrophoresis, Polyacrylamide Gel; Epitope; Epitope Mapping - methods; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Hemocyanin; Hemocyanins - chemistry; Hemocyanins - immunology; Molecular immunology; Panulirus interruptus; Techniques; PBS; BSA; Epitope; Hemocyanin; Fab; Competitive binding; Panulirus interruptus; Antigenic region; Antigenic determinant; Monoclonal antibody; Method; Macrura; Crustacea; Competitive fixation; Arthropoda; Decapoda; Invertebrata; Cuproprotein; Comparative study
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(95)00258-8

The competitive binding of a panel of monoclonal antibodies against hemocyanin of Panulirus interruptus hemocyanin was investigated with three different methods. A competitive-binding immunoassay method was more successful in the determination of ternary complexes than gel electrophoresis and gel filtration. The latter two methods could only be applied with antibodies possessing a higher affinity. Eleven monoclonal antibodies were assigned to groups on the basis of their interactions with five antigenic regions.