Enzyme-linked immunoassay. II. A simple method for synthesis of the rabbit antibody-beta-D-galactosidase complex and its general applicability.
Rabbit anti-human IgG antibody was mildly reduced in the presence of 10 mM 2-mercaptoethylamine and then coupled to beta-D-galactosidase [EC 3.2.1.23] from Escherichia coli using N,N'-o-phenylenedimaleimide. Human IgG and the anti-human IgG antibody-beta-D-galactosidase complex were successively adsorbed on Sepharose 4B binding rabbit anti-human IgG antibody. In this way amounts of human IgG as small as 5X10(-15) moles could be measured by determining the activity of beta-D-galactosidase bound to the Sepharose.
D000818:Animals; D000917:Antibody Formation; D005696:Galactosidases / Q000276:immunology; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000032:analysis; D011817:Rabbits / Q000276:immunology
D007074:Immunoglobulin G; D005696:Galactosidases