Aug 30, 1992

A monoclonal antibody-based enzyme immunoassay for human GMP-140/P-selectin

Journal of Immunological Methods
M KatayamaY Ikeda

Abstract

Two hybridoma cell lines producing monoclonal antibodies WGA-1 and PL7-6, reactive only with thrombin-stimulated human platelet have been established. Both these antibodies were investigated for their specific reactivity against GMP-140, based on the amino acid composition analysis of immunopurified antigen and N terminal amino acid sequencing of its protease fragments. A two-site enzyme immunoassay for quantification of human GMP-140 was developed using WGA-1 monoclonal antibody immobilized on 96-well microplates and horseradish peroxidase-labeled PL7-6 monoclonal antibody as detector. The assay was able to measure GMP-140 in serum and plasma with a sensitivity of about 5 ng/ml and a precision better than 10%. This assay will be useful for the detection of GMP-140 derived from platelets or endothelium in biological fluids and tissue extracts.

Mentioned in this Paper

Monoclonal Antibodies
SELP gene
Monoclonal antibodies, antineoplastic
Protein Sequence Determination
Mice, Inbred BALB C
F2 gene
Thrombin
Platelet Membrane Glycoproteins
Tissue Extracts
Endopeptidases

About this Paper

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