Addressing amino acid‐derived inhibitory metabolites and enhancing CHO cell culture performance through DOE‐guided media modifications

Author:

Ladiwala Pranay1ORCID,Dhara Venkata Gayatri1,Jenkins Jackson1,Kuang Bingyu2,Hoang Duc2ORCID,Yoon Seongkyu2ORCID,Betenbaugh Michael J.1ORCID

Affiliation:

1. Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore Maryland USA

2. Department of Chemical Engineering University of Massachusetts Lowell Lowell Massachusetts USA

Abstract

AbstractPreviously, we identified six inhibitory metabolites (IMs) accumulating in Chinese hamster ovary (CHO) cultures using AMBIC 1.0 community reference medium that negatively impacted culture performance. The goal of the current study was to modify the medium to control IM accumulation through design of experiments (DOE). Initial over‐supplementation of precursor amino acids (AAs) by 100% to 200% in the culture medium revealed positive correlations between initial AA concentrations and IM levels. A screening design identified 5 AA targets, Lys, Ile, Trp, Leu, Arg, as key contributors to IMs. Response surface design analysis was used to reduce initial AA levels between 13% and 33%, and these were then evaluated in batch and fed‐batch cultures. Lowering AAs in basal and feed medium and reducing feed rate from 10% to 5% reduced inhibitory metabolites HICA and NAP by up to 50%, MSA by 30%, and CMP by 15%. These reductions were accompanied by a 13% to 40% improvement in peak viable cell densities and 7% to 50% enhancement in IgG production in batch and fed‐batch processes, respectively. This study demonstrates the value of tuning specific AA levels in reference basal and feed media using statistical design methodologies to lower problematic IMs.

Funder

National Science Foundation

Publisher

Wiley

Subject

Applied Microbiology and Biotechnology,Bioengineering,Biotechnology

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