Document Detail

Monoclonal and polyclonal antibodies against porcine mitochondrial aspartate aminotransferase: their inhibition modes and application to enzyme immunoassay.
MedLine Citation:
PMID:  3297098     Owner:  NLM     Status:  MEDLINE    
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.
T Suzuki; Y Kishi; M Totani; H Kagamiyama; T Murachi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and applied biochemistry     Volume:  9     ISSN:  0885-4513     ISO Abbreviation:  Biotechnol. Appl. Biochem.     Publication Date:  1987 Apr 
Date Detail:
Created Date:  1987-07-24     Completed Date:  1987-07-24     Revised Date:  2007-03-21    
Medline Journal Info:
Nlm Unique ID:  8609465     Medline TA:  Biotechnol Appl Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  170-80     Citation Subset:  IM    
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MeSH Terms
Antibodies* / isolation & purification
Antibodies, Monoclonal* / isolation & purification
Aspartate Aminotransferases / analysis*,  immunology,  metabolism
Immunoenzyme Techniques
Mice, Inbred Strains
Mitochondria, Muscle / enzymology*
Reg. No./Substance:
0/Antibodies; 0/Antibodies, Monoclonal; EC Aminotransferases

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