Development and characterization of novel monoclonal antibodies against tartrate-resistant acid phosphatase 5

Tatsuya Ohashi, Toshihide Miura, Yoshihiko Igarashi, Iwao Kiyokawa, Yasuhito Sato, Kumiko Sasagawa, Ken Katagiri, Yoshiko Mochizuki, Takeshi Tomonaga, Fumio Nomura, Ryo Kojima, Katsuhiro Katayama

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Serum band 5 tartrate-resistant acid phosphatase (TRACP 5; EC 3.1.3.2) is a glycoprotein that exists as two very similar isoforms, TRACP 5a and TRACP 5b. The similarity of these two isoforins has made it difficult to establish monoclonal antibodies specific for either isoform. We report here the development of a monoclonal antibody with high specificity for TRACP 5b. We prepared TRACP 5b antigens from four sources: TRACP 5b purified from human bone, recombinant TRACP 5 from Escherichia coli, recombinant TRACP 5 from insect cells, and a synthetic TRACP 5b peptide. Thirty-seven mice were each immunized with 1 of the 4 different TRACP antigens to generate 473 antibody-producing clones. Three of these clones, Trk27, Trk49, and Trk62, reacted with TRACP 5b. These three clones were all established from mice exposed to native bone TRACP 5b antigen. In fact, none of the other antigens were able to generate anti-TRACP 5b monoclonal antibodies in mice. Furthermore, Trk62 interacted more strongly with TRACP 5b than with TRACP 5a. These results suggested that although recombinant proteins can be effective antigens, the native TRACP 5 protein might be more effective at generating monoclonal antibodies of greater specificity due to its more faithful representation of the native three-dimensional structure of the protein.

Original languageEnglish
Pages (from-to)358-366
Number of pages9
JournalHybridoma
Volume25
Issue number6
DOIs
Publication statusPublished - 2006 Dec
Externally publishedYes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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