Generation of Domain-Specific Monoclonal Antibodies Against Human Glutaredoxin3

Xin Dai, Yanqing Li, Xiaohong Sun, Kai Cai, Qinwen Mao, Haibin Xia*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Human Glutaredoxin3 (hGLRX3), which encodes a 37.4 kDa protein, possesses an N-terminal Trx homology domain followed by two tandem repeats of Grx domains. GLRX3 is expressed in many tissues and plays important roles in iron metabolism, antioxidant effect, cell proliferation and development, regulation of immune reaction, and tumorigenesis. The mechanisms underlying the biological function of GLRX3 are still not clear. To facilitate the functional research of GLRX3, in this study, monoclonal antibodies (MAbs) against hGLRX3 were produced by using purified prokaryotic recombinant 6His-hGLRX3 fusion protein as the immunogen. Five MAbs were obtained after preliminary screening by indirect enzyme-linked immunosorbent assay, then further characterized by Western blot analysis and immunocytochemistry. The domain specificity of these MAbs was also evaluated. Owing to the high conservation of protein sequences among different species, anti-GLRX3 MAbs produced in this study were shown to be immunoactive for GLRX3 in the cells from other species, such as mice, rats, Chinese hamster, and zebrafish. These domain-specific anti-GLRX3 MAbs will be an essential tool to investigate the roles of GLRX3 in normal physiological or pathological conditions.

Original languageEnglish (US)
Pages (from-to)285-292
Number of pages8
JournalMonoclonal Antibodies in Immunodiagnosis and Immunotherapy
Volume35
Issue number6
DOIs
StatePublished - Dec 2016

Keywords

  • Glutaredoxin3
  • hybridoma
  • monoclonal antibody
  • prokaryotic expression

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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