Acta Veterinaria et Zootechnica Sinica ›› 2024, Vol. 55 ›› Issue (12): 5762-5773.doi: 10.11843/j.issn.0366-6964.2024.12.038

• Basic Veterinary Medicine • Previous Articles     Next Articles

Preparation and Activity Analysis of Canine Derived Chimeric Single Chain Antibody scFv-Fc against Canine Distemper Virus

LIU Yakun1(), BI Zhenwei2,3,*(), XIA Xingxia2,3, MO Fei2,3, XU Siyu2,3, QIAN Jing2,3, TAN Yeping2,3, ZHU Yumei2,3, ZHAO Jianjun1,*()   

  1. 1. College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China
    2. Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
    3. GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China
  • Received:2024-02-04 Online:2024-12-23 Published:2024-12-27
  • Contact: BI Zhenwei, ZHAO Jianjun E-mail:910386496@qq.com;bizhenwei@126.com;zhaojianjun@byau.edu.cn

Abstract:

To reduce and improve the heterogeneity and efficacy of mouse-derived monoclonal antibodies in clinical treatment of dogs, the variable region of heavy chain (VH) and variable region of light chain (VL) were amplified from mouse-derived hybridomas secreting monoclonal antibody against canine distemper virus (CDV) using RT-PCR, the single chain antibody (scFv) gene was obtained by linking VH and VL through Linker, and the canine-derived chimeric single chain antibody (scFv-Fc) gene was obtained by selecting the constant region (Fc) of canine IgG B to connect to the C-terminus of scFv; The prokaryotic expression vectors for expressing mouse-derived scFv and canine-derived scFv-Fc were respectively constructed, which were expressed in Escherichia coli for purification, and the specificity and neutralizing activity of the expressed antibodies were detected by ELISA, IPMA, cell fusion inhibition, and neutralization assay. The detection limit of ELISA for CDV mouse-derived scFv and canine-derived scFv-Fc are 14.38 and 102.75 μg·mL-1, respectively; Both of scFv and scFv-Fc specifically react with the expressed CDV H protein and CDV; The cell membrane fusion mediated by CDV H and F proteins can be inhibited by scFv and scFv-Fc, and the minimum concentrations of them for complete protection of cells from CPE formation are 0.72 and 0.64 μg·mL-1, respectively. The mouse-derived scFv and canine-derived chimeric antibody scFv-Fc are successfully developed, providing candidate antibody drugs for the clinical treatment of canine distemper.

Key words: canine distemper virus, neutralizing activity, single chain antibody, chimeric antibody

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