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A quantitative enzyme-linked immunosorbent assay for rat insulin
1Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston 29425.
Journal of Immunoassay
|January 1, 1988
Summary
A new micro-Enzyme-Linked Immunosorbent Assay (ELISA) offers a sensitive and precise method for quantifying rat insulin. This assay demonstrates superior performance compared to radioimmunoassay (RIA), showing excellent accuracy and versatility across species.
Area of Science:
- Biochemistry
- Immunology
- Endocrinology
Background:
- Accurate quantification of insulin is crucial for understanding metabolic regulation and disease.
- Existing methods like radioimmunoassay (RIA) have limitations in sensitivity, range, or precision.
Purpose of the Study:
- To develop and validate a simple, quantitative micro-Enzyme-Linked Immunosorbent Assay (ELISA) for rat insulin.
- To compare the performance characteristics of the developed ELISA with a commercial RIA for rat insulin.
Main Methods:
- Development of a solid-phase, indirect, competitive micro-ELISA.
- Assay of pure rat insulin standards and insulin in pancreatic islet extracts.
- Comparison with a commercial rat insulin RIA, including range, sensitivity, accuracy, precision, and specificity.
Main Results:
- The micro-ELISA demonstrated a wider useful range (0.4-46.0 ng/ml) than RIA (0.2-8.6 ng/ml).
- ELISA sensitivity was 1.0 ng/ml (±0.13 ng/ml), favorably comparing to RIA (0.38 ±0.10 ng/ml).
- High correlation (r=0.91) with RIA in islet extracts; excellent linearity (r=0.97-0.99) with logit-log transformation; superior precision (intra-assay <10% CV, inter-assay ≤15% CV).
Conclusions:
- The developed micro-ELISA is a simple, quantitative, and versatile assay for rat insulin.
- It offers superior sensitivity, precision, and a wider assay range compared to commercial RIA.
- The assay is specific for rat insulin and adaptable for measuring insulin from other species, including human and porcine.