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Monoclonal antibodies expression improvement in CHO cells by PiggyBac transposition regarding vectors ratios and design

PLoS ONE. 2017-01; 
AhmadiSamira, DavamiFatemeh, DavoudiNoushin, NematpourFatemeh, AhmadiMaryam, EbadatSaeedeh, AzadmaneshKayhan, BarkhordariFarzaneh, MahboudiFerei
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Catalog Antibody … Human IgG (Genscript, Piscataway, USA) with defined concentration was applied to draw a standard curve. Antibody purification … Human standard IgG (Genscript, Piscataway, USA) was applied as the positive control in both SDS-PAGE and western blotting … Get A Quote

摘要

Establishing stable Chinese Hamster Ovary (CHO) cells producing monoclonal antibodies (mAbs) usually pass through the random integration of vectors to the cell genome, which is sensitive to gene silencing. One approach to overcome this issue is to target a highly transcribed region in the genome. Transposons are useful devices to target active parts of genomes, and PiggyBac (PB) transposon can be considered as a good option. In the present study, three PB transposon donor vectors containing both heavy and light chains were constructed, one contained independent expression cassettes while the others utilized either an Internal Ribosome Entry Site (IRES) or 2A element to express mAb. Conventional cell poo... More

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