title: Proteomic differences in recombinant CHO cells producing two similar antibody fragments.
abstract: 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.
authors.affiliation: 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.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Sommeregger
authors.affiliation: Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria.
authors.forename: Patrick
authors.initials: P
authors.lastname: Mayrhofer
authors.affiliation: Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria.
authors.forename: Willibald
authors.initials: W
authors.lastname: Steinfellner
authors.affiliation: Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 18, B, 5th Floor, 1190 Vienna, Austria.
authors.forename: David
authors.initials: D
authors.lastname: Reinhart
authors.affiliation: National Institute for Cellular Biotechnology (NICB), Dublin City University, Dublin 9, Ireland.
authors.forename: Michael
authors.initials: M
authors.lastname: Henry
authors.affiliation: National Institute for Cellular Biotechnology (NICB), Dublin City University, Dublin 9, Ireland.
authors.forename: Martin
authors.initials: M
authors.lastname: Clynes
authors.affiliation: National Institute for Cellular Biotechnology (NICB), Dublin City University, Dublin 9, Ireland. paula.meleady@dcu.ie.
authors.forename: Paula
authors.initials: P
authors.lastname: Meleady
authors.affiliation: 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.
authors.forename: Renate
authors.initials: R
authors.lastname: Kunert
mesh_terms: D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D018014:Gene Transfer Techniques; D008958:Models, Molecular; D055550:Protein Stability; D020543:Proteome / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000032:analysis / Q000378:metabolism*
chemical_list: D020543:Proteome; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies
keywords: Chinese hamster ovary cells; bottleneck; endoplasmic reticulum; secretion; specific productivity