Generation and characterization of a neutralizing rat anti-rmTNF-α monoclonal antibody

R. Lucas, K. Heirwegh, A. Neirynck, L. Remels, H. Van Heuverswyn, P. De Baetselier

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

A rat anti-recombinant mouse tumour necrosis factor-alpha (rmTNF-α) monoclonal IgM antibody (1F3F3) with high specific binding activity for rmTNF-α was generated. The 1F3F3 monoclonal antibody (mAb) neutralizes the cytotoxic activity in vitro of rmTNF-α on L929 cells and inhibits the binding of radiolabelled rmTNF-α to its putative receptor on L929 cells. The 1F3F3 mAb binds to monomeric, dimeric and trimeric rmTNF-α and does not bind to redued rmTNF-α, indicating that the recognized epitope is sensitive to denaturation. Using the 1F3F3 mAb as a capturing antibody and a biotinylated anti-rTNF-α as a detecting antibody, we have developed a sandwich ELISA that can specifically detect biologically active mTNF-α with a detection limit of 10 pg mTNF-α/well. This assay correlates well with the classical L929 cristal violet assay for the detection of bioactive rmTNF-α in biological fluids. The 1F3F3 mAb inhibits various in vitro biological activities of the rmTNF-α, such as the TNF-α-mediated tumouricidal activity of activated macrophages, the rmTNF-α-dependent stimulation of neutrophil degranulation and the growth-promoting effect of rmTNF-α. In vivo the 1F3F3 mAb inhibits lipopolysaccharide (LPS)-induced endotoxic shock. In conclusion, the 1F3F3 mAb is a useful tool to probe rmTNF-α activity both in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)218-223
Number of pages6
JournalImmunology
Volume71
Issue number2
StatePublished - Jan 1 1990
Externally publishedYes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Fingerprint Dive into the research topics of 'Generation and characterization of a neutralizing rat anti-rmTNF-α monoclonal antibody'. Together they form a unique fingerprint.

  • Cite this

    Lucas, R., Heirwegh, K., Neirynck, A., Remels, L., Van Heuverswyn, H., & De Baetselier, P. (1990). Generation and characterization of a neutralizing rat anti-rmTNF-α monoclonal antibody. Immunology, 71(2), 218-223.