Construction of mouse-derived phage display library of scFv fragments against Vibrio vulnificus VvhA

doi:10.3877/cma.j.issn.2095-9133.2021.05.007

Chinese Journal of Hygiene Rescue(Electronic Edition) ›› 2021, Vol. 07 ›› Issue (05): 287-291. doi: 10.3877/cma.j.issn.2095-9133.2021.05.007

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Construction of mouse-derived phage display library of scFv fragments against Vibrio vulnificus VvhA

Zhen Pan¹, Jiemin Chen¹, Chengjin Hu²^,^†()

^1. Department of Clinical Laboratory, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou 213000, China
^2. Department of Laboratory Medicine, 960 Hospital of PLA, Jinan 250031, China

Received:2021-06-22 Online:2021-10-18 Published:2021-12-14
Contact: Chengjin Hu

Abstract

Abstract:

Objective

To obtain highly specific VvhA cytolysin single-chain antibody, we used phage display technology to establish a murine phage single-chain antibody library.

Methods

After immunizing BALB/c mice with cytolysin protein, the total RNA was extracted from the spleen, and cDNA was obtained by reverse transcription. The cDNA was used as a template to amplify the antibody light chain and heavy chain variable region gene fragments, and OE-PCR technology was used to splice a complete scFv gene fragment. The recombinant phagemid pComb3XSS-scFv was constructed and electrotransformed into E. coli XL1-Blue to construct a phage single-chain antibody library.

Results

The recombination rate of the mouse-derived single-chain antibody phage library was 78% and the library capacity was 3×10⁸.

Conclusion

The murine single-chain antibody phage library is successfully constructed and lays the foundation for the early diagnosis of Vibrio vulnificus infection.

Key words: Vibrio vulnificus, VvhA, Phage display, scFv

Zhen Pan, Jiemin Chen, Chengjin Hu. Construction of mouse-derived phage display library of scFv fragments against Vibrio vulnificus VvhA[J]. Chinese Journal of Hygiene Rescue(Electronic Edition), 2021, 07(05): 287-291.

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Figures/Tables 9

References 14

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[1]	Yukun Guo, Yu′an Zhu, Xiaoming Fan, Feng Zhu, Bing Ma. Research progress on pathogenic mechanism of Vibrio vulnificus [J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2022, 17(01): 81-84.
[2]	Ruizhao Cai, Shaohai Qi. Biological characteristics and clinical research of Vibrio vulnificus [J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2020, 15(06): 490-494.

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