Frontiers in Immunology (May 2024)

Development of novel monoclonal antibodies for blocking NF-κB activation induced by CD2v protein in African swine fever virus

  • Rongrong Fan,
  • Rongrong Fan,
  • Zeliang Wei,
  • Zeliang Wei,
  • Zeliang Wei,
  • Mengmeng Zhang,
  • Shanshan Jia,
  • Zhiyang Jiang,
  • Yao Wang,
  • Junyu Cai,
  • Junyu Cai,
  • Guojiang Chen,
  • He Xiao,
  • Yinxiang Wei,
  • Yanchun Shi,
  • Jiannan Feng,
  • Beifen Shen,
  • Yuanqiang Zheng,
  • Yaojiang Huang,
  • Jing Wang

DOI
https://doi.org/10.3389/fimmu.2024.1352404
Journal volume & issue
Vol. 15

Abstract

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BackgroundCD2v, a critical outer envelope glycoprotein of the African swine fever virus (ASFV), plays a central role in the hemadsorption phenomenon during ASFV infection and is recognized as an essential immunoprotective protein. Monoclonal antibodies (mAbs) targeting CD2v have demonstrated promise in both diagnosing and combating African swine fever (ASF). The objective of this study was to develop specific monoclonal antibodies against CD2v.MethodsIn this investigation, Recombinant CD2v was expressed in eukaryotic cells, and murine mAbs were generated through meticulous screening and hybridoma cloning. Various techniques, including indirect enzyme-linked immunosorbent assay (ELISA), western blotting, immunofluorescence assay (IFA), and bio-layer interferometry (BLI), were employed to characterize the mAbs. Epitope mapping was conducted using truncation mutants and epitope peptide mapping.ResultsAn optimal antibody pair for a highly sensitive sandwich ELISA was identified, and the antigenic structures recognized by the mAbs were elucidated. Two linear epitopes highly conserved in ASFV genotype II strains, particularly in Chinese endemic strains, were identified, along with a unique glycosylated epitope. Three mAbs, 2B25, 3G25, and 8G1, effectively blocked CD2v-induced NF-κB activation.ConclusionsThis study provides valuable insights into the antigenic structure of ASFV CD2v. The mAbs obtained in this study hold great potential for use in the development of ASF diagnostic strategies, and the identified epitopes may contribute to vaccine development against ASFV.

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