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Title: IRES-mediated Tricistronic vectors for enhancing generation of high monoclonal antibody expressing CHO cell lines
Authors: Ho, S.C.L.
Bardor, M.
Feng, H.
Tong, Y.W. 
Song, Z.
Yap, M.G.S. 
Yang, Y.
Keywords: Aggregation
Monoclonal antibody
Issue Date: Jan-2012
Citation: Ho, S.C.L., Bardor, M., Feng, H., Mariati, Tong, Y.W., Song, Z., Yap, M.G.S., Yang, Y. (2012-01). IRES-mediated Tricistronic vectors for enhancing generation of high monoclonal antibody expressing CHO cell lines. Journal of Biotechnology 157 (1) : 130-139. ScholarBank@NUS Repository.
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. © 2011 Elsevier B.V.
Source Title: Journal of Biotechnology
ISSN: 01681656
DOI: 10.1016/j.jbiotec.2011.09.023
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