Homology modelling and bivalent single-chain Fv construction of anti-HepG2 single-chain immunoglobulin Fv fragments from a phage display library
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- 作者：Ming Ni ; Bing Yu ; Yu Huang ; Zhenjie Tang ; Ping Lei ; Xin Shen…
- 关键词：BsFv ; homology modelling ; scFv ; three ; dimensional (3D) structure
- 刊名：Journal of Biosciences
- 出版年：2008
- 出版时间：December 2008
- 年：2008
- 卷：33
- 期：5
- 页码：691-697
- 全文大小：
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Bing Yu (1)
Yu Huang (2)
Zhenjie Tang (2)
Ping Lei (2)
Xin Shen (2)
Wei Xin (2)
Huifen Zhu (2)
Guanxin Shen (2)
3. Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China
1. Department of Pathogen Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China
2. Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China - 刊物类别：Life Sciences, general; Biomedicine general; Zoology; Plant Sciences; Microbiology; Cell Biology;
- 刊物主题：Life Sciences, general; Biomedicine general; Zoology; Plant Sciences; Microbiology; Cell Biology;
- 出版者：Springer India
- ISSN：0973-7138
文摘
We prepared single-chain immunoglobulin Fv fragments (scFv) SLH10 specific for the HepG2 cell line after biopanning from a large human-na茂e 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. Keywords BsFv homology modelling scFv three-dimensional (3D) structure