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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Virology
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1459402

Expression and characterization of recombinant antibodies against H7 subtype avian influenza virus and their diagnostic potential

Provisionally accepted
Siwen Wang Siwen Wang 1Ying Zhang Ying Zhang 2Xu Zhou Xu Zhou 1Yue Ma Yue Ma 1Jianzhong Shi Jianzhong Shi 1Yongping Jiang Yongping Jiang 1Yanbing Li Yanbing Li 1Xiurong Wang Xiurong Wang 1*
  • 1 Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
  • 2 College of Life Engineering, Shenyang Institute of Technology, Fushun, China

The final, formatted version of the article will be published soon.

    Monoclonal antibodies (mAbs) play a pivotal role in disease diagnosis as well as immunotherapy interventions. Traditional monoclonal antibody generation relies on animal immunization procedures predominantly involving mice; however, recent advances in in-vitro expression methodologies have enabled large-scale production suitable for both industrial applications as well as scientific investigations. In this study, two mAbs against H7 subtype avian influenza viruses (AIV) were sequenced and analyzed, and the DNA sequences encoding heavy chain (HC) and light chain (LC) were obtained and cloned into pCHO-1.0 expression vector. Then, the HC and LC expression plasmids were transfected into CHO-S cells to establish stable cell lines expressing these mAbs using a twophase selection scheme with different concentrations of methotrexate and puromycin. Recombinant antibodies were purified from the cell culture medium, and their potential applications were evaluated using hemagglutination inhibition (HI), western blotting (WB), confocal microscopy, and enzymelinked immunosorbent assay (ELISA). The results indicated that the obtained recombinant antibodies exhibited biological activity similar to that of the parent antibodies derived from ascites and could be used as a replacement for animal-derived mAbs. A kinetic analysis of the two antibodies to the AIV HA protein, conducted using surface plasmon resonance (SPR), showed concordance between the recombinant and parental antibodies. The data presented in this study suggest that the described antibody production protocol could avoid the use of experimental animals and better conform to animal welfare regulations, and provides a basis for further research and development of mAbs-based diagnostic products.

    Keywords: avian influenza viruses, H7 subtype, recombinant antibody, monoclonal antibodies, CHO-s

    Received: 04 Jul 2024; Accepted: 24 Jul 2024.

    Copyright: © 2024 Wang, Zhang, Zhou, Ma, Shi, Jiang, Li and Wang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Xiurong Wang, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.