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Proteins, Expression, Isolation and Analysis> | … cytometry (FCM). 2.2. Expression and purification of 5T4 ECD-Fc. The extracellular domain of human 5T4 (residues 32-355) was fused into the C-terminal human IgG Fc tag to construct 5T4-ECD-Fc (Genscript, China). FreeStyle … | Get A Quote |
5T4, a membrane protein, is overexpressed in many tumor tissues but rarely expressed in normal tissues. Here, CHO-5T4 cells were generated and served as the antigen to immunize mice. Hybridoma techniques were employed to produce monoclonal antibodies (mAbs). The recombinant protein of human IgG Fc-fused extracellular domain of 5T4 (5T4 ECD-Fc) was obtained from transient expression in HEK293F cells. The fusion protein 5T4 ECD-Fc and CHO-5T4 cells were respectively utilized to screen anti-5T4 antibodies that could bind to the native antigen. In preliminary screening, three hundred and fifty mAbs were obtained. Via surface plasmon resonance and flow cytometry screening, seven anti-5T4 mAbs stood out. Am... More