An avidin-biotin ELISA for the measurement of mitochondrial aspartate aminotransferase in human serum.
We describe a new double-antibody ELISA system using avidin-biotin amplication for the mass measurement of mitochondrial aspartate aminotransferase (m-AST) in human serum. The assay is very sensitive and as little as 0.5 ng/ml of m-AST may be detected. The method is linear up to 100 micrograms/l. The within-day and day-to-day coefficients of variation were found to be 8.9% and 11.5% respectively for a low m-AST concentration (2.7 micrograms/l), and 5.9% and 8.0% respectively for a high level of m-AST (30 micrograms/l). The assay requires 100 microliters of serum and can be completed within 5 h. The ELISA procedure and a classical immunoprecipitation technique measuring the catalytic activity of the isoenzyme were applied simultaneously to the sera of 189 subjects. The protein levels determined by ELISA correlated poorly (r = 0.66) with the catalytic activity of m-AST.
Varasteh
A
A
Centre du Médicament, UA CNRS 597, Nancy, France.
Wellman
M
M
Artur
Y
Y
Schiele
F
F
Siest
G
G
D000293:Adolescent; D000918:Antibody Specificity; D001219:Aspartate Aminotransferases / Q000097:blood* / Q000276:immunology / Q000378:metabolism; D001360:Avidin; D001710:Biotin; D002384:Catalysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007527:Isoenzymes / Q000097:blood / Q000378:metabolism; D008297:Male; D008928:Mitochondria / Q000201:enzymology*; D011233:Precipitin Tests; D015203:Reproducibility of Results
D007527:Isoenzymes; D001360:Avidin; D001710:Biotin; D001219:Aspartate Aminotransferases