title: A monoclonal antibody-based enzyme immunoassay for human GMP-140/P-selectin.
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.
authors.affiliation: Biotechnology Research Laboratories, Takara Shuzo Co., Ltd, Shiga, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Katayama
authors.forename: M
authors.initials: M
authors.lastname: Handa
authors.forename: H
authors.initials: H
authors.lastname: Ambo
authors.forename: Y
authors.initials: Y
authors.lastname: Araki
authors.forename: S
authors.initials: S
authors.lastname: Hirai
authors.forename: I
authors.initials: I
authors.lastname: Kato
authors.forename: Y
authors.initials: Y
authors.lastname: Kawai
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
authors.forename: Y
authors.initials: Y
authors.lastname: Ikeda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins