Screening and preparation of a human single chain antibody(scFv)against Rabies Virus G protein from a phage display library and identification of the neutralizing activity
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    Abstract:

    Objective:To construct a human scFv phage antibody library against rabies virus G protein,selected the single chain antibody against rabies virus G protein for different epitopes,and identificated its neutralizing activity. Methods:The human scFv phage antibody library against rabies virus G protein was constructed by using peripheral blood lymphocytes from 130 Rabies vaccine immune volunteers. Screening by the purified rabies virus G protein,the positive clones were identified and expressed,identified neutralizing activity by AMMS. Results:Through the prokaryotic expression system and purification of rabies virus G protein,the human anti rabies virus G protein scFv phage antibody library was constructed with about 5.0×107 size. The nucleic acid sequence analysis confirmed the insertion of a fragment is scFv. After 5 rounds of screening,we randomly picked 140 clones by phage-ELISA,and obtained 4 nucleic acid sequences in different scFv antibodies,purified and identified activity. We finally obtained 1 scFv antibody with neutralizing activity. Conclusion:We have successfully constructed a whole human anti-rabies virus G protein scFv antibody library,and obtained a whole human anti-rabies virus G protein scFv antibody with neutralizing activity from the library. It is established foundation of the new type of rabies virus vaccine for human immunization.

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张倩倩,赵 茜,张 晓,丁贵鹏,金 秋,高 畅,熊四平,陈玉平,朱 进,冯振卿.全人源抗狂犬病病毒G蛋白单链抗体制备及中和活性鉴定[J].南京医科大学学报(自然科学版英文版),2013,(7):927-931.

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  • Received:February 13,2013
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  • Online: July 09,2013
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