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Overexpression of Serpinb1 in Chinese hamster ovary cells increases recombinant IgG productivity

Paper ID Volume ID Publish Year Pages File Format Full-Text
22974 43408 2015 9 PDF Available
Title
Overexpression of Serpinb1 in Chinese hamster ovary cells increases recombinant IgG productivity
Abstract

•First report on a novel CHO cell engineering target, Serpinb1, for improving IgG expression.•The lentiviral pool overexpressing SERPINB1 increased 1.3 fold in VCD compared with control.•Peak IgG volumetric productivity increased 2.0 fold in this lentiviral pool compared with control.•This work represents ongoing applications of “-omics” findings to CHO host cell line engineering.

We report the discovery and validation of a novel CHO cell engineering target for improving IgG expression, serpin peptidase inhibitor, clade B, member 1 (Serpinb1). Transcriptomic studies using microarrays revealed that Serpinb1 was up-regulated in cultures with IgG heavy and light chain transcription transiently repressed compared with cultures treated with non-targeting siRNA. As proof of concept, a lentiviral vector was employed to overexpress the Chinese Hamster Serpinb1 in a CHOZN® Glutamine Synthetase (−/−) recombinant IgG producing CHO line. The lentiviral stable pool demonstrated 4.2-fold SERPINB1 overexpression compared with the non-transduced control. The peak viable cell density (VCD) and peak IgG volumetric productivity of the lentiviral stable pool increased 1.3 and 2.0 fold, respectively, compared with the non-transduced control. For host cell engineering, a plasmid encoding SERPINB1 was transfected into the CHOZN® GS (−/−) host cell line to create several stable pools. Single-cell clones isolated from the pools were characterized for their SERPINB1 expression levels and growth. The clone (SERPINB1_OE_27) with the highest SERPINB1 expression had decreased peak viable cell density and exponential phase growth rate. Selected SERPINB1 OE clones were subsequently evaluated for their IgG expression capabilities using GS selection. Clone SERPINB1_OE_42 with moderate SERPINB1 overexpression demonstrated increased IgG productivity in “bulk” selection. We conclude that manipulating Serpinb1 expression can lead to increased recombinant IgG productivity, but the effect in host cell lines may vary by clone and by overexpression level. This work represents the ongoing effort in applying “-omics” findings to novel CHO host cell line engineering.

Keywords
Chinese hamster ovary cells; Host cell engineering; Serpinb1; Overexpression; Productivity
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Overexpression of Serpinb1 in Chinese hamster ovary cells increases recombinant IgG productivity
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 193, 10 January 2015, Pages 91–99
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