A non-competitive solid-phase radioimmunoassay for human aldolase A.
A solid-phase, non-competitive radioimmunoassay for aldolase A in human serum has been developed. Human aldolase A was purified from muscle, and specific antisera to the purified aldolase A were obtained from chickens. Specific IgG anti-human aldolase A was purified by affinity chromatography. Disposable polypropylene plates were coated with specific IgG antibody and used for radioimmunoassay with 125I-specific IgG antibody to aldolase A. The non-specific binding was minimized by saturating the binding sites of the plates with 2% ovalbumin in 0.1% Tween 20. This radioimmunoassay is specific for the aldolase A subunit, with no cross-reactivity with human aldolase B subunit or homopolymeric human aldolase C(C4). The serum aldolase A immunoreactivities of 33 normal subjects ranged from 124 to 212 ng/ml with a mean of 178 +/- 41 ng/ml (+/- 2 SD). Ninety-three patients' sera were assayed with both a solid-phase non-competitive radioimmunoassay and a competitive double antibody radioimmunoassay developed in our laboratory and the results showed a high degree of correlation (r = 0.912; p less than 0.001). Rapidity and simplicity of the solid-phase assay makes it superior to other methods for the measurement of serum aldolase isozymes.
D000818:Animals; D000918:Antibody Specificity; D002645:Chickens / Q000276:immunology; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis*; D007700:Kinetics; D011863:Radioimmunoassay / Q000379:methods; D013997:Time Factors; D014020:Tissue Extracts / Q000032:analysis
D014020:Tissue Extracts; D005634:Fructose-Bisphosphate Aldolase