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Expression and purification of recombinant antibody

QZ Qi Zhang
MG Miao Gui
XN Xuefeng Niu
SH Shihua He
RW Ruoke Wang
YF Yupeng Feng
AK Andrea Kroeker
YZ Yanan Zuo
HW Hua Wang
YW Ying Wang
JL Jiade Li
CL Chufang Li
YS Yi Shi
XS Xuanling Shi
GG George F. Gao
YX Ye Xiang
XQ Xiangguo Qiu
LC Ling Chen
LZ Linqi Zhang
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The PCR-amplified and sequence-confirmed the variable regions of heavy and light chain genes were separately cloned into backbone of antibody expression vectors containing the constant regions of human IgG160. Whole recombinant human IgG1 antibodies were expressed in 293T cells (ATCC) by transient transfection and purified by affinity chromatography using Protein A agarose. The concentration was determined by BCA Protein Assay Kit (Thermo Scientific). The expression clones encoding the control antibody KZ52, c13C6, mAb114 were synthesized and confirmed by sequencing before production.

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