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      https://www.ias.ac.in/article/fulltext/jbsc/033/05/0691-0697

    • Keywords

       

      BsFv; homology modelling; scFv; three-dimensional (3D) structure

    • Abstract

       

      We prepared single-chain immunoglobulin Fv fragments (scFv) SLH10 specific for the HepG2 cell line after biopanning from a large human-naïve phage display library (Griffin. 1 Library). The three-dimensional (3D) structure of SLH10 was modelled by the Insight II molecule simulation software. The structure was refined using the molecular dynamics method. The structures with the least steric clashes and lowest energy were determined finally. The optimized structures of heavy (VH) and light (VL) variable chains of SLH10 scFv were obtained. Then SLH10 bivalent single-chain Fv (BsFv) was constructed that would be suitable for high-affinity targeting. SLH10 BsFv was generated by linking scFvs together and identified by sequencing. Its expression products were confirmed by western blot analysis. The relative molecular masses of scFv and BsFv were approximately 30 kDa and 60 kDa, respectively. Flow cytometry revealed that SLH10 BsFv bound the selected cell lines with greater signal intensity than the parental scFv. The improved antigen binding of SLH10 BsFv may be useful for immunodiagnostics or targeted gene therapy for liver cancer.

    • Author Affiliations

       

      Ming Ni1 Bing Yu2 Y U Huang3 Zhenjie Tang3 Ping Lei3 Xin Shen3 Wei Xin3 Huifen Zhu3 Guanxin Shen3

      1. Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
      2. Department of Pathogen Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
      3. Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
    • Dates

       
  • Journal of Biosciences | News

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