Multiplex Secretome Engineering Enhances Recombinant Protein Production and Purity

Overview

Journal Nat Commun

Specialty Biology

Date 2020 Apr 22

PMID 32313013

Citations 37

Authors

Stefan Kol

Daniel Ley

Tune Wulff

Marianne Decker

Johnny Arnsdorf

Sanne Schoffelen

Anders Holmgaard Hansen

Tanja Lyholm Jensen

Jahir M Gutierrez

Austin W T Chiang

Helen O Masson

Bernhard O Palsson

Bjorn G Voldborg

Lasse Ebdrup Pedersen

Helene Faustrup Kildegaard

Gyun Min Lee

Nathan E Lewis

Affiliations

Soon will be listed here.

Abstract

Host cell proteins (HCPs) are process-related impurities generated during biotherapeutic protein production. HCPs can be problematic if they pose a significant metabolic demand, degrade product quality, or contaminate the final product. Here, we present an effort to create a "clean" Chinese hamster ovary (CHO) cell by disrupting multiple genes to eliminate HCPs. Using a model of CHO cell protein secretion, we predict that the elimination of unnecessary HCPs could have a non-negligible impact on protein production. We analyze the HCP content of 6-protein, 11-protein, and 14-protein knockout clones. These cell lines exhibit a substantial reduction in total HCP content (40%-70%). We also observe higher productivity and improved growth characteristics in specific clones. The reduced HCP content facilitates purification of a monoclonal antibody. Thus, substantial improvements can be made in protein titer and purity through large-scale HCP deletion, providing an avenue to increased quality and affordability of high-value biopharmaceuticals.

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