A gel electrophoresis method for epitope mapping studies with monoclonal antibodies.
The linear polyacrylamide gradient gel electrophoresis method of Lambin and Fine (1979, Anal. Biochem. 98, 160-168) has been adapted for estimation of the molecular weights of antigen-antibody complexes, thereby providing information useful in epitope mapping studies with monoclonal antibodies. The method has been applied to mapping of the epitopes recognized by four different monoclonal antibodies raised against rat brain hexokinase (1984, Finney et al., J. Biol. Chem., 259, 8232-8237). The results obtained with the gel electrophoresis method were in agreement with epitope mapping studies conducted by the molecular sieve high-performance liquid chromatographic method of Crawford et al., (1982, Proc. Natl. Acad. Sci. USA 79, 7031-7035). Epitope mapping by the gel electrophoretic method offers several advantages in terms of speed, convenience, and economy when compared with alternative procedures that have been described.
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D001666:Binding Sites, Antibody; D001921:Brain / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis*; D006593:Hexokinase / Q000276:immunology; D007120:Immunochemistry; D008970:Molecular Weight; D051381:Rats
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000939:Epitopes; D006593:Hexokinase