High-Throughput Method for Screening Recombinant Coagulation Factor VII Expressing Cells

doi:10.3969/j.issn.1673-1689.2018.07.008

Home > Archive>Volume 37, Issue 7, 2018 >722-726. DOI:10.3969/j.issn.1673-1689.2018.07.008

High-Throughput Method for Screening Recombinant Coagulation Factor VII Expressing Cells
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                        10.3969/j.issn.1673-1689.2018.07.008
                    
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Abstract:

To overcome the time-consuming screening process，mammalian cells expressing recombinant therapeutic protein were selected by flow cytometry and method of cell purity analysis was established. Transfected CHO cells were sorted by flow cytometer and high expression sub-clone was screened by dot blot and western blot. CHO cells after transfecting with rFVII expression vector were divided by flow cytometry to 96 micro-plates and recombinant coagulation factor VII expression was analyzed by dot blot and western blot. High rFVII expression clone，CHO-rFVII-1，was obtained and the purity of CHO-rFVII-1 was 99.9 %. Moreover，CHO-rFVII-1 had similar viable cell density and rFVII expression compared to CHO-rFVII，which was obtained by traditional screening method. Screening high expression clones by flow cytometry was a rapid and high-throughput method and will be beneficial to screen high recombinant therapeutic protein expression clone in other mammalian cells.

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PENG Lin, LI Chengyuan, XIONG Wendian, CAI Yanfei, JIN Jian, LI Huazhong. High-Throughput Method for Screening Recombinant Coagulation Factor VII Expressing Cells[J]. Journal of Food Science and Biotechnology,2018,37(7):722-726.

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Online: September 28,2018
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