Module-Specific Antibodies Against Human Connective Tissue Growth Factor: Utility for Structural and Functional Analysis of the Factor as Related to Chondrocytes

Masanao Minato, Satoshi Kubota, Harumi Kawaki, Takashi Nishida, Akira Miyauchi, Hiroshi Hanagata, Tohru Nakanishi, Teruko Takano-Yamamoto, Masaharu Takigawa

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Connective tissue growth factor/hypertrophic chondrocyte specific gene product 24 (CTGF/Hcs24/CCN2) shows diverse functions in the process of endochondral ossification. It promotes not only the proliferation and differentiation of chondrocytes and osteoblasts in vitro, but also angiogenesis in vivo. The ctgf gene is a member of the gene family called CCN, and it encodes the characteristic 4-module structure of this family, with the protein containing IGFBP, VWC, TSP and CT modules. We raised several monoclonal antibodies and polyclonal antisera against CTGF, and located the epitopes in the modules by Western blotting. For mapping the epitopes, Brevibacillus-produced independent modules were utilized. As a result, at least 1 antibody or antiserum was prepared for the detection of each module in CTGF. Western blotting with these antibodies is expected to be useful for the analysis of CTGF fragmentation. Moreover, we examined the effects of these monoclonal antibodies on the biological functions of CTGF. One out of 3 humanized monoclonal antibodies was found to neutralize efficiently the stimulatory effect of CTGF on chondrocytic cell proliferation. This particular antibody bound to the CT module. In contrast, surprisingly, all of the 3 antibodies recognizing IGFBP, VWC and CT modules stimulated proteoglycan synthesis in chondrocytic cells. Together with previous findings, these results provide insight into the structural-functional relationships of CTGF in executing multiple functions.

Original languageEnglish
Pages (from-to)347-354
Number of pages8
JournalJournal of biochemistry
Volume135
Issue number3
DOIs
Publication statusPublished - 2004 Mar
Externally publishedYes

Keywords

  • CCN family
  • Chondrocyte
  • Connective tissue growth factor
  • Module
  • Monoclonal antibody

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Minato, M., Kubota, S., Kawaki, H., Nishida, T., Miyauchi, A., Hanagata, H., Nakanishi, T., Takano-Yamamoto, T., & Takigawa, M. (2004). Module-Specific Antibodies Against Human Connective Tissue Growth Factor: Utility for Structural and Functional Analysis of the Factor as Related to Chondrocytes. Journal of biochemistry, 135(3), 347-354. https://doi.org/10.1093/jb/mvh042