PMID- 37096798
OWN - NLM
STAT- MEDLINE
DCOM- 20230817
LR  - 20230817
IS  - 1097-0290 (Electronic)
IS  - 0006-3592 (Linking)
VI  - 120
IP  - 9
DP  - 2023 Sep
TI  - Addressing amino acid-derived inhibitory metabolites and enhancing CHO cell 
      culture performance through DOE-guided media modifications.
PG  - 2542-2558
LID - 10.1002/bit.28403 [doi]
AB  - Previously, 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.
CI  - (c) 2023 The Authors. Biotechnology and Bioengineering published by Wiley 
      Periodicals LLC.
FAU - Ladiwala, Pranay
AU  - Ladiwala P
AUID- ORCID: 0000-0002-6231-254X
AD  - Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 
      Baltimore, Maryland, USA.
FAU - Dhara, Venkata Gayatri
AU  - Dhara VG
AD  - Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 
      Baltimore, Maryland, USA.
FAU - Jenkins, Jackson
AU  - Jenkins J
AD  - Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 
      Baltimore, Maryland, USA.
FAU - Kuang, Bingyu
AU  - Kuang B
AD  - Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, 
      Massachusetts, USA.
FAU - Hoang, Duc
AU  - Hoang D
AUID- ORCID: 0000-0002-7496-5193
AD  - Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, 
      Massachusetts, USA.
FAU - Yoon, Seongkyu
AU  - Yoon S
AUID- ORCID: 0000-0002-5330-8784
AD  - Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, 
      Massachusetts, USA.
FAU - Betenbaugh, Michael J
AU  - Betenbaugh MJ
AUID- ORCID: 0000-0002-6336-4659
AD  - Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 
      Baltimore, Maryland, USA.
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20230425
PL  - United States
TA  - Biotechnol Bioeng
JT  - Biotechnology and bioengineering
JID - 7502021
RN  - 0 (Amino Acids)
RN  - 0 (Culture Media)
SB  - IM
MH  - Cricetinae
MH  - Animals
MH  - Cricetulus
MH  - *Amino Acids/metabolism
MH  - CHO Cells
MH  - Culture Media/chemistry
MH  - *Batch Cell Culture Techniques/methods
OTO - NOTNLM
OT  - Chinese hamster ovary
OT  - amino acid metabolism
OT  - design of experiments
OT  - fed-batch bioprocessing
OT  - inhibitory metabolites
OT  - medium optimization
EDAT- 2023/04/25 17:42
MHDA- 2023/08/17 06:43
CRDT- 2023/04/25 07:02
PHST- 2023/03/22 00:00 [revised]
PHST- 2022/12/18 00:00 [received]
PHST- 2023/04/10 00:00 [accepted]
PHST- 2023/08/17 06:43 [medline]
PHST- 2023/04/25 17:42 [pubmed]
PHST- 2023/04/25 07:02 [entrez]
AID - 10.1002/bit.28403 [doi]
PST - ppublish
SO  - Biotechnol Bioeng. 2023 Sep;120(9):2542-2558. doi: 10.1002/bit.28403. Epub 2023 
      Apr 25.