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Enzyme linked immunosorbent assay for measuring antithrombin III.
P Edgar1, I Jennings, P Harper
1Department of Haematology, Addenbrooke's Hospital, Cambridge.
Journal of Clinical Pathology
|September 1, 1989
Summary
A new competitive ELISA assay was developed to measure antithrombin III antigen. This sensitive and precise immunoassay shows strong correlation with existing methods for antithrombin III measurement.
Area of Science:
- Biochemistry
- Immunology
- Clinical Chemistry
Background:
- Antithrombin III is a crucial protein in the blood coagulation cascade.
- Accurate measurement of antithrombin III antigen is important for diagnosing and managing thrombotic disorders.
Purpose of the Study:
- To develop and validate a novel competitive enzyme-linked immunosorbent assay (ELISA) for quantifying antithrombin III antigen.
- To assess the sensitivity, precision, and correlation of the developed ELISA with a standard method.
Main Methods:
- Development of a competitive ELISA using purified antithrombin III.
- A two-step immunoassay protocol minimizing non-specific antibody binding.
- Validation using plasma samples compared against immune electrophoresis.
Main Results:
- The developed ELISA demonstrated high sensitivity and precision.
- Plasma samples measured by ELISA showed excellent correlation with immune electrophoresis (r = 0.933).
- The assay design avoids direct binding of anti-antithrombin antibody to the solid phase.
Conclusions:
- The competitive ELISA is a reliable and accurate method for measuring antithrombin III antigen.
- This assay offers a sensitive and precise alternative for clinical laboratory use.
- The assay's performance suggests its utility in diagnosing and monitoring conditions related to antithrombin III levels.