用单克隆抗体检测小鼠骨桥蛋白的各种表位

Detection of various epitopes of murine osteopontin by monoclonal antibodies.

作者信息

Hotta H, Kon S, Katagiri Y U, Tosa N, Tsukamoto T, Chambers A F, Uede T

机构信息

Institute of Immunological Science, Hokkaido University, Kita-15, Niishi-7, Kita-ku, Sapporo, Japan.

出版信息

Biochem Biophys Res Commun. 1999 Apr 2;257(1):6-11. doi: 10.1006/bbrc.1999.0412.

DOI:10.1006/bbrc.1999.0412

PMID:10092501

Abstract

We immunized rats with recombinant murine osteopontin protein and obtained four monoclonal antibodies recognizing distinct epitopes of murine osteopontin. OPN1.2 recognized the amino-terminal half of OPN, while OPN2.2, OPN2.3, and OPN3.1 recognized the carboxy-terminal half of OPN. The epitope recognized by OPN2.2 was destroyed by further cleavage of the carboxy half of OPN. The epitope recognized by OPN2.3 was located in the amino-terminal end of the carboxy half of OPN, whereas that recognized by OPN3.1 was located in the carboxy-terminal end of the carboxy half of OPN. OPN1.2 and OPN2.2 recognized thrombin-cleaved osteopontin, whereas thrombin-cleaved osteopontin was not recognized by OPN2.3 and OPN3.1. Thus, these monoclonal antibodies will be useful in structure/function studies of the role of osteopontin in murine models of disease.

摘要

我们用重组鼠骨桥蛋白免疫大鼠，获得了四种识别鼠骨桥蛋白不同表位的单克隆抗体。OPN1.2识别骨桥蛋白的氨基端一半区域，而OPN2.2、OPN2.3和OPN3.1识别骨桥蛋白的羧基端一半区域。OPN2.2识别的表位在骨桥蛋白羧基端一半区域进一步切割后被破坏。OPN2.3识别的表位位于骨桥蛋白羧基端一半区域的氨基端，而OPN3.1识别的表位位于骨桥蛋白羧基端一半区域的羧基端。OPN1.2和OPN2.2识别凝血酶切割后的骨桥蛋白，而OPN2.3和OPN3.1不识别凝血酶切割后的骨桥蛋白。因此，这些单克隆抗体将有助于在鼠疾病模型中研究骨桥蛋白作用的结构/功能研究。