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Degradation of recombinant proteins by CHO host cell proteases is prevented by Matriptase-1 knock-out

Laux, Holger and Romand, Sandrine and Nuciforo, Sandro and Farady, Christopher and Tapparel, Joel Alois Rene and Buechmann-Moeller, Stine and Sommer, Benjamin and Oakeley, Edward James and Bodendorf, Ursula (2018) Degradation of recombinant proteins by CHO host cell proteases is prevented by Matriptase-1 knock-out. Biotechnology and bioengineering. ISSN 00063592

Abstract

An increasing number of non-antibody format proteins are entering the clinical development. However, one of the major hurdles for the production of non-antibody glycoproteins is host cell-related proteolytic degradation, which can drastically impact developability and timelines of pipeline projects.
Chinese hamster ovary (CHO) cells are the preferred production host for recombinant therapeutic proteins. Using protease inhibitors, transcriptomics and genetic knockdowns we have identified, out of the more than 700 known proteases in rodents, Matriptase-1 as the major protease involved in degradation of recombinant proteins expressed in CHO-K1 cells. Subsequently Matriptase-1 was deleted in CHO-K1 cells using “Transcription Activator-Like Effector Nucleases” (TALENs) as well as zinc-finger nucleases (ZFNs). This resulted in a superior CHO-K1 matriptase knockout (KO) cell line with strongly reduced or no proteolytic degradation activity towards a panel of recombinantly-expressed proteins.
The matriptase KO cell line was evaluated in spike-in experiments, and showed little or no degradation of proteins incubated in conditioned medium derived from the KO cells. This effect was confirmed when the same proteins were recombinantly expressed in the KO cell line.
In summary, the combination of novel cell line engineering tools, next generation sequencing screening methods and the recently published Chinese hamster genome has enabled the development of this novel matriptase KO CHO cell line capable of improving expression yields of intact therapeutic proteins.

Item Type: Article
Date Deposited: 19 Jun 2018 00:45
Last Modified: 19 Jun 2018 00:45
URI: https://oak.novartis.com/id/eprint/36047

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