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Construction and transient expression of the mouse-human chimeric antibody against human CD28

Cheng-Gang; Chen-Yong-Jing; Ma-Zhen-Yu; Fei-Min; Sun-Zhong-Wen; Xu-Yao-Yu; Hu-Ling-Ling; Qiu-Yu-Hua

Fudan Xuebao 34(4): 482-485

2007

ISSN/ISBN: 1672-8467
Accession: 015357969
Purpose To construct and transiently express of a mouse-human chimeric antibody against human CD28 molecule.Methods By using genetic technology, total RNA was extracted from the murine hybridoma cell line (Clone number: 2F5) secreting anti-CD28 monoclonal antibody. The variable region and the correspond signal peptide coding sequence were cloned, then fused with the coding sequence of the constant region of the human IgG1 heavy chain and light chain respectively. The mouse-human chimeric antibody recombinant plasmid was established. 293T cell line was transfected with the recombinant plasmid and the transfectants were selected by G418 contained medium. Using FACS to detect the chimeric antibody's transient expression level.Results The chimeric antibody against human CD28 was expressed and recognized the CD28 molecular specifically.Conclusions Chimeric antibody is successfully constructed and is transiently expressed, thus a foundation is laid for establishing CHO cell line secreting chimeric anti-CD28 monoclonal antibody.

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