Proteomic differences in recombinant CHO cells producing two similar antibody fragments.

Autor: Sommeregger W; Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria.; Polymun Scientific GmbH, Klosterneuburg, Austria.; Bilfinger Industrietechnik Salzburg GmbH, Salzburg, Austria., Mayrhofer P; Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria., Steinfellner W; Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria., Reinhart D; Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria., Henry M; National Institute for Cellular Biotechnology (NICB), Dublin City University, Dublin 9, Ireland., Clynes M; National Institute for Cellular Biotechnology (NICB), Dublin City University, Dublin 9, Ireland., Meleady P; National Institute for Cellular Biotechnology (NICB), Dublin City University, Dublin 9, Ireland. paula.meleady@dcu.ie., Kunert R; Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria. renate.kunert@boku.ac.at.
Jazyk: angličtina
Zdroj: Biotechnology and bioengineering [Biotechnol Bioeng] 2016 Sep; Vol. 113 (9), pp. 1902-12. Date of Electronic Publication: 2016 Mar 16.
DOI: 10.1002/bit.25957
Abstrakt: Chinese hamster ovary (CHO) cells are the most commonly used mammalian hosts for the production of biopharmaceuticals. To overcome unfavorable features of CHO cells, a lot of effort is put into cell engineering to improve phenotype. "Omics" studies investigating elevated growth rate and specific productivities as well as extracellular stimulus have already revealed many interesting engineering targets. However, it remains largely unknown how physicochemical properties of the recombinant product itself influence the host cell. In this study, we used quantitative label-free LC-MS proteomic analyses to investigate product-specific proteome differences in CHO cells producing two similar antibody fragments. We established recombinant CHO cells producing the two antibodies, 3D6 and 2F5, both as single-chain Fv-Fc homodimeric antibody fragments (scFv-Fc). We applied three different vector strategies for transgene delivery (i.e., plasmid, bacterial artificial chromosome, recombinase-mediated cassette exchange), selected two best performing clones from transgene variants and transgene delivery methods and investigated three consecutively passaged cell samples by label-free proteomic analysis. LC-MS-MS profiles were compared in several sample combinations to gain insights into different aspects of proteomic changes caused by overexpression of two different heterologous proteins. This study suggests that not only the levels of specific product secretion but the product itself has a large impact on the proteome of the cell. Biotechnol. Bioeng. 2016;113: 1902-1912. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.
(© 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.)
Databáze: MEDLINE
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