title: Monoclonal and polyclonal antibodies against porcine mitochondrial aspartate aminotransferase: their inhibition modes and application to enzyme immunoassay.
abstract: Monoclonal and polyclonal antibodies against porcine mitochondrial aspartate aminotransferase (m-AST) were prepared in order to study their effect on the kinetics of the enzyme and their possible use as diagnostic reagents. The most stable hybridoma clone, designated MH-1, was selected and cultured for mass production of the monoclonal antibody MA-1. MA-1 was purified by affinity chromatography with m-AST as a ligand. The m-AST activity was inhibited uncompetitively by preincubation with MA-1, but preincubation with the polyclonal antibody uncompetitively by preincubation with MA-1, but preincubation with the polyclonal antibody raised in a rabbit resulted in noncompetitive inhibition of the enzyme. These results suggest the usefulness of a monoclonal antibody for studying the mechanism of catalysis. Sandwich enzyme immunoassay methods for m-AST using both polyclonal and monoclonal antibody-coated polystyrene balls were established and permitted the determination of porcine m-AST on the order of 10(-11) and 10(-10)M, respectively.
authors.forename: T
authors.initials: T
authors.lastname: Suzuki
authors.forename: Y
authors.initials: Y
authors.lastname: Kishi
authors.forename: M
authors.initials: M
authors.lastname: Totani
authors.forename: H
authors.initials: H
authors.lastname: Kagamiyama
authors.forename: T
authors.initials: T
authors.lastname: Murachi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001219:Aspartate Aminotransferases / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D007124:Immunoenzyme Techniques; D007700:Kinetics; D051379:Mice; D008815:Mice, Inbred Strains; D008931:Mitochondria, Muscle / Q000201:enzymology*; D013552:Swine
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D001219:Aspartate Aminotransferases