Type

Journal Article

Authors

Renate Kunert
Paula Meleady
Martin Clynes
Michael Henry
David Reinhart
Willibald Steinfellner
Patrick Mayrhofer
Wolfgang Sommeregger

Subjects

Biochemistry

Topics
growth rate antibody producing cells cho cells single chain fv recombinant antibody fragments chinese hamster ovary cells engineering physicochemical properties

Proteomic differences in recombinant CHO cells producing two similar antibody fragments. (2015)

Abstract Chinese hamster ovary (CHO) cells are the most commonly used mammalian hosts for the production of biopharmaceuticals. To overcome unfavourable 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. This article is protected by copyright. All rights reserved.
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Full list of authors on original publication

Renate Kunert, Paula Meleady, Martin Clynes, Michael Henry, David Reinhart, Willibald Steinfellner, Patrick Mayrhofer, Wolfgang Sommeregger

Experts in our system

1
Paula Meleady
Dublin City University
Total Publications: 95
 
2
Martin Clynes
Dublin City University
Total Publications: 209
 
3
Michael Henry
Dublin City University
Total Publications: 78