A new monoclonal antibody, mAb 204-11, that influences the binding of platelet GPVI to fibrous collagen

Masaaki Moroi; Jun Mizuguchi; Sachiko Kawashima; Michiko Nagamatsu; Yoshiki Miura; Tomohiro Nakagaki; Katsuaki Ito; Stephanie M. Jung

doi:10.1055/s-0037-1613401

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2003; 89(06): 996-1003
DOI: 10.1055/s-0037-1613401

Platelets and Blood Cells

Schattauer GmbH

A new monoclonal antibody, mAb 204-11, that influences the binding of platelet GPVI to fibrous collagen

Masaaki Moroi

¹Department of Protein Biochemistry, Institute of Life Science, Kurume University, Kurume, Fukuoka, Japan

,

Jun Mizuguchi

²Blood Products Research Department, The Chemo-Sero-Therapeutic Research Institute, Kumamoto, Kumamoto, Japan

,

Sachiko Kawashima

³Department of Veterinary Pharmacology, Faculty of Agriculture, Miyazaki University, Miyazaki, Miyazaki, Japan

,

Michiko Nagamatsu

²Blood Products Research Department, The Chemo-Sero-Therapeutic Research Institute, Kumamoto, Kumamoto, Japan

,

Yoshiki Miura

¹Department of Protein Biochemistry, Institute of Life Science, Kurume University, Kurume, Fukuoka, Japan

,

Tomohiro Nakagaki

²Blood Products Research Department, The Chemo-Sero-Therapeutic Research Institute, Kumamoto, Kumamoto, Japan

,

Katsuaki Ito

³Department of Veterinary Pharmacology, Faculty of Agriculture, Miyazaki University, Miyazaki, Miyazaki, Japan

,

Stephanie M. Jung

¹Department of Protein Biochemistry, Institute of Life Science, Kurume University, Kurume, Fukuoka, Japan

› Institutsangaben

Abstract

Summary

The newly identified platelet collagen receptor glycoprotein VI binds to fibrous collagen, inducing platelet activation. Several antibodies against GPVI have been reported, including a patient’s auto-antibodies, that activates platelets through their ability to crosslink this glycoprotein. We have developed a monoclonal antibody (mAb) against GPVI using the recombinant extracellular domain of GPVI as an antigen. This antibody, mAb 204-11, induced platelet aggregation and tyrosine phosphorylation of proteins similar to those induced by GPVI-reactive proteins, collagen and convulxin. Its interaction with GPVI was analyzed by measuring the effect of the antibody on GPVI binding to collagen using a dimeric form of recombinant GPVI, GPVI-Fc₂. MAb 204-11 inhibited the binding of GPVI-Fc₂ to fibrous collagen particles, but enhanced the GPVI binding to immobilized collagen, suggesting that the antibody binds to a region near the collagen binding site of GPVI. MAb 204-11 also inhibited the GPVI binding to convulxin at a low concentration, but not completely. Since mAb 204-11 reacts specifically with GPVI and is applicable for immunoblotting and immunoprecipitation, this antibody would be useful for studies on GPVI.

Keywords

Platelet - GPVI - monoclonal antibody - collagen receptor - collagen

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Referenzen

References
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