Four-way ligation for construction of a mammalian cell-based full-length antibody display library |
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Authors: | Zhou Ivan Zhang Zhe-Huan Li Chang-Zheng Chen Zhen-Rui He Wei Zhou Ye Liu Shuwen Wu Shuguang Zhou Yuanping Tan Wanlong Jiang Shibo Zhou Chen |
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Institution: | Antiviral Research Center, School of Pharmaceutical Science, Southern Medical University, Guangzhou, China. |
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Abstract: | A unique four-way ligation strategy was developed for rapid construction of a full-length antibody library. A mammalian expression vector was constructed that contained dual mammalian expression cassettes and sequences recognized by the unique restriction enzymes BsmBI, BstXI, and SfiI. Both full-length light-chain and variable domain of heavy-chain genes were inserted into the vector in one step by four-way ligation, and full-length bivalent antibodies were displayed on mammalian cell surfaces. Using this strategy, only 2 weeks were required to successfully construct high-quality, full-length human antibody libraries. |
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