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IRES-mediated Tricistronic vectors for enhancing generation of high monoclonal antibody expressing CHO cell lines

Abstract : A Tricistronic vector utilizing internal ribosome entry site (IRES) elements to express the light chain (LC), heavy chain (HC), and a neomycin phosphotransferase (NPT) selection marker from one transcript is designed for generation of mAb expressing CHO cell lines. As compared to the commonly used vectors, benefits of this design include: (1) minimized non-expressing clones, (2) enhanced stable mAb productivity without gene amplification, (3) control of LC and HC expression at defined ratios, and (4) consistent product quality. After optimization of the LC and HC arrangement and increasing selection stringency by weakening the NPT selection marker, this Tricistronic vector is able to generate stably transfected pools with specific productivity (qmAb) greater than 5 pg/cell/day (pcd) and titers over 150 mg/L. 5% of clones from these pools have qmAb greater than 20 pcd and titers ranging from 300 to more than 500 mg/L under non-optimized shake flask batch cultures using commercially available protein-free medium. The mAb produced by these clones have low aggregation and consistent glycosylation profiles. The entire process of transfection to high-expressing clones requires only 6 months. The IRES-mediated Tricistronic vector provides an attractive alternative to commonly used vectors for fast generation of mAb CHO cell lines with high productivity.
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https://hal-normandie-univ.archives-ouvertes.fr/hal-01844673
Contributor : Arnaud Lehner <>
Submitted on : Thursday, July 19, 2018 - 3:44:21 PM
Last modification on : Friday, April 5, 2019 - 3:24:07 PM

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Steven C.L. Ho, Muriel Bardor, Huatao Feng, . Mariati, Yen Wah Tong, et al.. IRES-mediated Tricistronic vectors for enhancing generation of high monoclonal antibody expressing CHO cell lines. African Journal of Biotechnology, Academic Journals, 2012, 157 (1), pp.130 - 139. ⟨10.1016/j.jbiotec.2011.09.023⟩. ⟨hal-01844673⟩

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