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A non-competitive solid-phase radioimmunoassay for human aldolase A
Clinica Chimica Acta; International Journal of Clinical Chemistry
|October 13, 1982
Summary
A new radioimmunoassay accurately measures aldolase A in human serum. This rapid and simple method offers a superior alternative for quantifying aldolase isozymes.
Area of Science:
- Biochemistry
- Immunology
- Clinical Chemistry
Background:
- Aldolase A is a key enzyme in glycolysis.
- Accurate measurement of serum aldolase A is important for diagnosing certain conditions.
- Existing methods for aldolase isozyme measurement can be complex or lack specificity.
Purpose of the Study:
- To develop a solid-phase, non-competitive radioimmunoassay for human serum aldolase A.
- To establish a specific, sensitive, and reproducible assay for aldolase A quantification.
Main Methods:
- Purification of human aldolase A from muscle tissue.
- Generation of specific chicken antisera against aldolase A.
- Affinity chromatography for IgG purification.
- Coating of polypropylene plates with IgG antibody.
- Use of 125I-labeled IgG antibody for detection.
- Minimization of non-specific binding using ovalbumin and Tween 20.
Main Results:
- The developed radioimmunoassay demonstrated high specificity for aldolase A, with no cross-reactivity to aldolase B or C.
- Serum aldolase A levels in normal subjects ranged from 124 to 212 ng/ml (mean 178 +/- 41 ng/ml).
- Results from the solid-phase assay highly correlated (r = 0.912) with a competitive double antibody radioimmunoassay.
Conclusions:
- A novel, solid-phase, non-competitive radioimmunoassay for serum aldolase A has been successfully developed.
- The assay is specific, reproducible, and provides accurate quantification of aldolase A.
- The rapidity and simplicity of this method make it advantageous for measuring serum aldolase isozymes compared to existing techniques.