alexa Proteomic Analysis of Bioreactor Cultures of an Antibody Expressing CHOGS Cell Line that Promotes High Productivity
ISSN: 0974-276X

Journal of Proteomics & Bioinformatics
Open Access

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Research Article

Proteomic Analysis of Bioreactor Cultures of an Antibody Expressing CHOGS Cell Line that Promotes High Productivity

Haimanti Dorai1#*, Suli Liu3#, Xiang Yao2, Yonghui Wang1, Uelke Tekindemir1, Michael J Lewis1, Shiaw-Lin Wu3 and William Hancock3

1Janssen Research and Development, Welsh and McKean Rd, Springhouse, PA 19477, USA

2Janssen Research and Development, 3210 Merryfield Row, San Diego, CA 92121, USA

3Barnett Institute of Chemical and Biological Analysis, 140 The Fenway, Boston, MA 02115, USA

#These two authors contributed equally

*Corresponding Author:
Haimanti Dorai
Janssen Research and Development
Welsh and McKean Rd
Springhouse, PA 19477, USA
E-mail: [email protected]; or Shiaw-Lin Wu: [email protected]

Received date: April 17, 2013; Accepted date: May 21, 2013; Published date: May 25, 2013

Citation: Dorai H, Liu S, Yao X, Wang Y, Tekindemir U, et al. (2013) Proteomic Analysis of Bioreactor Cultures of an Antibody Expressing CHO-GS Cell Line that Promotes High Productivity. J Proteomics Bioinform 6: 099-108. doi: 10.4172/jpb.1000268

Copyright: © 2013 Dorai H, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Antibody manufacturing cell line development at Janssen Research & Development involves transfection of therapeutic antibody genes into a CHO-GS host cell line and isolating primary transfectomas that upon cloning yield high expressing cell lines secreting the desired antibody products. Subsequently, these cell lines are cultivated in stirred tank bioreactors for the large-scale generation of the products. In an attempt to optimize this process for high productivity, a two pronged approach was undertaken.

First, in a Design of Experiment study, a CHO-GS cell line expressing a therapeutic antibody was cultivated in 2 L DasGip fed-batch mini-bioreactors under a variety of culture conditions. In general, culture conditions that promoted robust growth and high viable cell density resulted in high productivity. Then, cell culture harvests and cell lysates from two ‘high productivity’ and two ‘low productivity’ bioreactors were subjected to proteomic analysis using the CHO genome database, on two independent days. The levels of each protein expressed in these two sets of bioreactors were then compared. A total of 180 proteins that were modulated two-fold or more were thus identified, only 12 of which were consistently modulated across multiple days in culture. The modulated proteins have biological process functions that are related to cytoskeleton rearrangement, protein synthesis, cell metabolism and cell growth. Provided that these observations are validated by Western blot, one or more of these proteins, whose expression correlated to productivity can potentially be utilized as targets for manipulating a superior transfection host cell line. At a minimum, the expression levels of these proteins can provide insight for further process optimization efforts.

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