title: Thermostabilization of carboxypeptidase A by interaction with its monoclonal antibodies.
abstract: The effect of interaction of carboxypeptidase A (CPA) with three monoclonal antibodies, each with a different epitope (CP 10, CP 9, and CP 8), on the heat stabilization of enzymes is described. These monoclonal antibodies bind to CPA with a relatively high binding constant (approximately 10(8) M-1) and do not affect its catalytic properties. Intact carboxypeptidase A lost more than 95 and 90% of its esterase and peptidase activities within 120 min at 50 degrees C. The monoclonal antibodies increased the thermal stability of the enzyme by 90 and 60%, as compared with the peptidase and esterase initial activities, respectively. Binding of these monoclonal antibodies, alone or in pairs, to the enzyme epitopes that are supposedly involved in heat denaturation of CPA result in stabilization of the conformation of the enzyme. The effect of thermostabilization by monoclonal antibodies was more pronounced with respect to peptidase activity than to esterase activity, indicating that these activities follow different reaction mechanisms. Since properly selected monoclonal antibodies can be prepared against virtually any enzyme, their immunocomplexation may provide a general and convenient method for stabilization of the enzyme conformation to heat denaturation, without affecting the catalytic properties.
authors.affiliation: Department of Biotechnology and Microbiology, Faculty of Life Sciences, Tel-Aviv University, Israel.
authors.forename: B
authors.initials: B
authors.lastname: Solomon
authors.forename: N
authors.initials: N
authors.lastname: Balas
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism*; D002268:Carboxypeptidases / Q000276:immunology / Q000378:metabolism*; D043422:Carboxypeptidases A; D004795:Enzyme Stability; D004797:Enzyme-Linked Immunosorbent Assay; D004950:Esterases / Q000378:metabolism; D007700:Kinetics; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D004950:Esterases; D002268:Carboxypeptidases; D043422:Carboxypeptidases A