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An in-house ELISA method for quantification of circulating tissue factor

Tay, S.P., and Cheong, S.K., (2002) An in-house ELISA method for quantification of circulating tissue factor. Malaysian Journal of Pathology, 24 (1). pp. 45-51. ISSN 0126-8635

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Official URL: http://www.mjpath.org.my/past_issue/MJP2002.1/An%20in-house%20ELISA%20method%20for%20quantification%20of%20circulating%20tissue%20factor.pdf

Affiliations

Universiti Kebangsaan Malaysia, Malaysia, Faculty of Medicine, Dept. of Pathology

Abstract

Tissue Factor (TF) is a low molecular weight transmembrane glycoprotein that initiates the clotting protease cascade. It is considered to be the principal regulator of the extrinsic coagulation pathway, hemostasis and thrombosis, as well as inflammation and cellular immune response. An in-house two-step direct sandwich ELISA (enzyme-linked immunosorbent assay) for immunological quantification of plasma TF was successfully developed. The assay employed a monoclonal antibody against human TF (1:400 dilution; 1250 nglml) and peroxidase-conjugated anti-TF IgG(1:1000 dilution; 2000 ng/ml) as capture and detecting antibodies respectively, whilst tetramethylbenzidine/H202 were utilized as substrates. Titration curves of recombinant TF were linear within 10 to 4000 pglml, with a detection limit of 36.3 1 pglml. It demonstrated low intra- (2.50 - 9.23 CV%) and inter-assays (5.65 - 13.57 CV%) variability, as well as satisfactory analytical recovery (91.55 - 103.95%) and good parallelism. The assay developed was intended to be applied for measurement of plasma TF levels in patients with thrombotic disorders.

Item Type:Journal
Additional Information:The financial support under the IRPA grant No. 06-02-02-0067 from the Ministry of Science, Technology and Environment is gratefully acknowledged.
Keywords:Tissue Factor, enzyme-linked immunosorbent assay, thrombotic disorders
Subjects:R Medicine, Dentistry, Pharmacy, Nursing
ID Code:1823

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