ePrints@IIScePrints@IISc Home | About | Browse | Latest Additions | Advanced Search | Contact | Help

Development and Characterization of a Potent Tumor Necrosis Factor-Alpha-Blocking Agent

Jameei, A and Nagarajan, D and Sarikhani, M and Chandra, N and Karande, AA (2019) Development and Characterization of a Potent Tumor Necrosis Factor-Alpha-Blocking Agent. In: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, 38 (4). pp. 145-156.

mon_ant_imm_imm_38-4_145-156_2019.pdf - Published Version

Download (954kB) | Preview
Official URL: https://doi.org/10.1089/mab.2019.0018


Tumor necrosis factor-α (TNFα), one of the major proinflammatory cytokines, plays a key role in an effective immune response. However, the chronic presence of TNFα can lead to several inflammatory disorders, such as rheumatoid arthritis, psoriasis, Crohn's disease, etc. Inhibition of TNFα by pharmacological inhibitors or antibodies has proven to be effective in palliative treatment to some extent. The aim of this study was to develop an anti-TNFα antibody, which may be used as a therapeutic option to inhibit TNFα-mediated cytotoxicity. We characterized several hybridoma clones secreting monoclonal antibodies (mAbs) to human-TNFα. Four mAbs rescued L929 fibroblast cells from TNFα-triggered cell death and one of these, namely C8, was found to have the highest affinity. To gain insights into the mechanism by which mAb C8 inhibits human TNFα-mediated toxicity, the epitope corresponding to the mAb was delineated. The antigenic determinant was found to comprise of the stretch of amino acids 99-120, of which, 102-104 (glutamine, arginine, glutamic acid) form the core epitope. The observation was supported by bioinformatics analyses of an antigen/antibody complex model. In addition, the binding affinity of mAb C8 to TNFα was found to be comparable with that of infliximab, which is a commercially available anti-TNFα mAb.

Item Type: Journal Article
Publication: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Publisher: Mary Ann Liebert Inc.
Additional Information: The copyright for this article belongs to the Authors.
Keywords: arginine; epitope; glutamic acid; glutamine; infliximab; monoclonal antibody; monoclonal antibody C8; polyhistidine tag; tumor necrosis factor; tumor necrosis factor inhibitor; unclassified drug; immunoglobulin G; monoclonal antibody; recombinant protein; tumor necrosis factor, animal cell; antagonist potency; Article; binding affinity; bioinformatics; cell death; clone; conformational transition; controlled study; cytotoxicity; drug binding site; drug mechanism; drug receptor binding; enzyme linked immunosorbent assay; enzyme phosphorylation; expression vector; female; human; hybridoma; immunoblotting; molecular docking; mouse; nonhuman; priority journal; animal; antibody production; Bagg albino mouse; biosynthesis; cell culture; cytology; fibroblast; immunology, Animals; Antibodies, Monoclonal, Humanized; Antibody Formation; Cells, Cultured; Female; Fibroblasts; Humans; Hybridomas; Immunoglobulin G; Mice; Mice, Inbred BALB C; Recombinant Proteins; Tumor Necrosis Factor-alpha
Department/Centre: Division of Biological Sciences > Biochemistry
Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 21 Oct 2022 11:06
Last Modified: 21 Oct 2022 11:06
URI: https://eprints.iisc.ac.in/id/eprint/77484

Actions (login required)

View Item View Item