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Rapid production of a chimeric antibody-antigen fusion protein based on 2A-peptide cleavage and green fluorescent protein expression in CHO cells

MAbs.. 2019-01; 
Van der Weken H, Cox E, Devriendt B
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Gene Synthesis A sequence coding for a recombinant chimeric mouse-porcine IgA-FedF fusion antibody, separated by furin + GT2A peptide sequences was synthesized by GenScript and cloned into the pcDNA3.1- N-eGFP vector at the KpnI restriction site... Get A Quote
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摘要

To enable large-scale antibody production, the creation of a stable, high producer cell line is essential. This process often takes longer than 6 months using standard limited dilution techniques and is very labor intensive. The use of a tri-cistronic vector expressing green fluorescent protein (GFP) and both antibody chains, separated by a GT2A peptide sequence, allows expression of all proteins under a single promotor in equimolar ratios. By combining the advantages of 2A peptide cleavage and single cell sorting, a chimeric antibody-antigen fusion protein that contained the variable domains of mouse IgG with a porcine IgA constant domain fused to the FedF antigen could be produced in CHO-K1 cells. After trans... More

关键词

2A peptide; CHO; FACS; GFP; antibody engineering; chimeric; expression; production; recombinant antibody