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Background <p>Feline calicivirus (FCV) is a significant causative pathogen of upper respiratory tract disease and virulent systemic infection in feline populations, yet specific targeted antiviral agents for FCV infection remain unavailable.</p> Methods <p>A feline-derived single-chain variable fragment (scFv) phage display library targeting the VP1-CDE neutralizing region of FCV was constructed using peripheral blood mononuclear cells isolated from cats immunized with recombinant VP1-CDE protein. Positive scFv clones were screened via three rounds of affinity panning and phage ELISA. The scFv-Fc fusion proteins were expressed in eukaryotic cells, and their antigen-binding properties and antiviral potential were characterized through Western blotting, bio-layer interferometry, immunofluorescence assay and viral neutralization assay.</p> Results <p>The constructed library reached a size of 2.1 × 10<sup>8</sup> independent phages with favorable sequence diversity. Five unique scFv clones, designated 3A5, 3B8, 3H5, 2H5, and 2I5, were identified. All five clones could specifically bind to both recombinant VP1-CDE protein and FCV-infected F81 feline kidney cells. Viral neutralization assays demonstrated that scFv-Fc-3A5 exhibited the most potent neutralizing activity against the genogroup II FCV LZ2023 isolate (IC<sub>50</sub>= 1.862&#xa0;µg/mL), scFv-Fc-2H5 showed weak neutralization (IC<sub>50</sub>= 6.168&#xa0;µg/mL), and the remaining three clones had no appreciable neutralizing capacity.</p> Conclusions <p>scFv-Fc-3A5 and scFv-Fc-2H5 are promising candidate molecules for the development of diagnostic reagents and antiviral therapeutics targeting genogroup II FCV.</p>

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Isolation of feline-derived scFvs against the VP1-CDE region of feline calicivirus from a phage display library and characterization of their antigen-binding and antiviral potential in vitro

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月27日

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  • Yanquan Wei,
  • Hongmei Wang,
  • Zhen Han,
  • Huitian Gou,
  • Dongdong Sun,
  • Yihan Liu

摘要

Background

Feline calicivirus (FCV) is a significant causative pathogen of upper respiratory tract disease and virulent systemic infection in feline populations, yet specific targeted antiviral agents for FCV infection remain unavailable.

Methods

A feline-derived single-chain variable fragment (scFv) phage display library targeting the VP1-CDE neutralizing region of FCV was constructed using peripheral blood mononuclear cells isolated from cats immunized with recombinant VP1-CDE protein. Positive scFv clones were screened via three rounds of affinity panning and phage ELISA. The scFv-Fc fusion proteins were expressed in eukaryotic cells, and their antigen-binding properties and antiviral potential were characterized through Western blotting, bio-layer interferometry, immunofluorescence assay and viral neutralization assay.

Results

The constructed library reached a size of 2.1 × 108 independent phages with favorable sequence diversity. Five unique scFv clones, designated 3A5, 3B8, 3H5, 2H5, and 2I5, were identified. All five clones could specifically bind to both recombinant VP1-CDE protein and FCV-infected F81 feline kidney cells. Viral neutralization assays demonstrated that scFv-Fc-3A5 exhibited the most potent neutralizing activity against the genogroup II FCV LZ2023 isolate (IC50= 1.862 µg/mL), scFv-Fc-2H5 showed weak neutralization (IC50= 6.168 µg/mL), and the remaining three clones had no appreciable neutralizing capacity.

Conclusions

scFv-Fc-3A5 and scFv-Fc-2H5 are promising candidate molecules for the development of diagnostic reagents and antiviral therapeutics targeting genogroup II FCV.