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Optimization of cell line development in the GS-CHO expression system using a high-throughput, single cell-based clone selection system

Paper ID Volume ID Publish Year Pages File Format Full-Text
20196 43162 2015 7 PDF Available
Title
Optimization of cell line development in the GS-CHO expression system using a high-throughput, single cell-based clone selection system
Abstract

•We applied the high-throughput ClonePix FL for cell line development using CHO-K1SV.•We conducted optimization of the semi-solid medium formulation for CHO-K1SV.•We showed the importance of spent medium and recombinant Albumine in semi-solid media.•The cell culture duration after transfection affected the colony formation ratio.•Considerations of optimized conditions are seemed to be useful for other cell lines.

Therapeutic antibodies are commonly produced by high-expressing, clonal and recombinant Chinese hamster ovary (CHO) cell lines. Currently, CHO cells dominate as a commercial production host because of their ease of use, established regulatory track record, and safety profile. CHO-K1SV is a suspension, protein-free-adapted CHO-K1-derived cell line employing the glutamine synthetase (GS) gene expression system (GS-CHO expression system). The selection of high-producing mammalian cell lines is a crucial step in process development for the production of therapeutic antibodies. In general, cloning by the limiting dilution method is used to isolate high-producing monoclonal CHO cells. However, the limiting dilution method is time consuming and has a low probability of monoclonality. To minimize the duration and increase the probability of obtaining high-producing clones with high monoclonality, an automated single cell-based clone selector, the ClonePix FL system, is available. In this study, we applied the high-throughput ClonePix FL system for cell line development using CHO-K1SV cells and investigated efficient conditions for single cell-based clone selection. CHO-K1SV cell growth at the pre-picking stage was improved by optimizing the formulation of semi-solid medium. The efficiency of picking and cell growth at the post-picking stage was improved by optimization of the plating time without decreasing the diversity of clones. The conditions for selection, including the medium formulation, were the most important factors for the single cell-based clone selection system to construct a high-producing CHO cell line.

Keywords
Chinese hamster ovary cell; Glutamine synthetase; Cell line development; High-throughput selection; Semi-solid media
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Optimization of cell line development in the GS-CHO expression system using a high-throughput, single cell-based clone selection system
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 120, Issue 3, September 2015, Pages 323–329
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us