Four-way ligation for construction of a mammalian cell-based full-length antibody display library.
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.
Antiviral Research Center, School of Pharmaceutical Science, Southern Medical University, Guangzhou, China.
Ivan
I
Zhou
Zhe-Huan
ZH
Zhang
Chang-Zheng
CZ
Li
Zhen-Rui
ZR
Chen
Wei
W
He
Ye
Y
Zhou
Shuwen
S
Liu
Shuguang
S
Wu
Yuanping
Y
Zhou
Wanlong
W
Tan
Shibo
S
Jiang
Chen
C
Zhou
D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence / Q000235:genetics; D002460:Cell Line; D006224:Cricetinae; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D014162:Transfection / Q000379:methods*
D000906:Antibodies