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A new method for the estimation of protein C by ELISA
Thrombosis Research
|December 15, 1984
Summary
A new enzyme-linked immunosorbent assay (ELISA) accurately quantifies human protein C antigen. This method offers a sensitive and specific alternative to electroimmunodiffusion for assessing protein C levels in various patient groups.
Area of Science:
- Biochemistry
- Immunology
- Clinical Chemistry
Background:
- Human protein C is a crucial vitamin K-dependent plasma glycoprotein involved in regulating blood coagulation.
- Accurate quantitation of protein C antigen is essential for diagnosing and managing bleeding disorders and thrombosis.
- Existing methods like electroimmunodiffusion (EID) may have limitations in sensitivity or throughput.
Purpose of the Study:
- To develop and validate a novel solid-phase enzyme-linked immunosorbent assay (ELISA) for quantifying human protein C antigen.
- To compare the performance of the developed ELISA with the established Laurell technique (EID).
- To assess protein C antigen levels in various clinical conditions, including congenital deficiency, liver cirrhosis, and dicoumarol treatment.
Main Methods:
- Development of a solid-phase ELISA using anti-protein C F(ab')2 fragments and peroxidase-labeled anti-protein C IgG.
- Quantification of protein C antigen in plasma samples from healthy controls and patients with specific conditions.
- Comparison of ELISA results with those obtained by EID for correlation analysis.
Main Results:
- The developed ELISA demonstrated high specificity, sensitivity (0.02% detection limit), and accuracy (3-10% variation coefficient).
- Excellent correlation (r=0.95) was observed between ELISA and EID results across all tested plasma samples.
- Significantly lower protein C levels were found in liver cirrhosis patients compared to other vitamin K-dependent factors. In dicoumarol-treated patients, protein C decreased more rapidly than Factors X or II.
Conclusions:
- The novel ELISA is a reliable, sensitive, and accurate method for quantifying human protein C antigen.
- This ELISA provides a valuable tool for clinical diagnostics and research, offering advantages over traditional methods like EID.
- The study highlights the utility of the ELISA in characterizing protein C levels in diverse pathological states and during anticoagulant therapy.