PMID- 36892270
OWN - NLM
STAT- MEDLINE
DCOM- 20230613
LR  - 20230613
IS  - 1860-7314 (Electronic)
IS  - 1860-6768 (Linking)
VI  - 18
IP  - 6
DP  - 2023 Jun
TI  - Cottonseed hydrolysate supplementation alters metabolic and proteomics responses 
      in Chinese hamster ovary cell cultures.
PG  - e2200243
LID - 10.1002/biot.202200243 [doi]
AB  - Hydrolysates are used as media supplements although their role is not well 
      characterized. In this study, cottonseed hydrolysates, which contained peptides 
      and galactose as supplemental substrates, were added to Chinese hamster ovary 
      (CHO) batch cultures, enhancing cell growth, immunoglobulin (IgG) titers, and 
      productivities. Extracellular metabolomics coupled with tandem mass tag (TMT) 
      proteomics revealed metabolic and proteomic changes in cottonseed-supplemented 
      cultures. Shifts in production and consumption dynamics of glucose, glutamine, 
      lactate, pyruvate, serine, glycine, glutamate, and aspartate suggest changes in 
      tricarboxylic acid (TCA) and glycolysis metabolism following hydrolysate inputs. 
      Quantitative proteomics revealed 5521 proteins and numerous changes in relative 
      abundance of proteins related to growth, metabolism, oxidative stress, protein 
      productivity, and apoptosis/cell death at day 5 and day 6. Differential abundance 
      of amino acid transporter proteins and catabolism enzymes such as 
      branched-chain-amino-acid aminotransferase (BCAT)1 and fumarylacetoacetase (FAH) 
      can alter availability and utilization of several amino acids. Also, pathways 
      involved in growth including the polyamine biosynthesis through higher ornithine 
      decarboxylase (ODC1) abundance and hippo signaling were upregulated and 
      downregulated, respectively. Central metabolism rewiring was indicated by 
      glyceraldehyde-3-phosphate dehydrogenase (GAPDH) downregulation, which 
      corresponded with re-uptake of secreted lactate in the cottonseed-supplemented 
      cultures. Overall, cottonseed hydrolysate supplementation modified culture 
      performance by altering cellular activities critical to growth and protein 
      productivity including metabolism, transport, mitosis, transcription, 
      translation, protein processing, and apoptosis. HIGHLIGHTS: Cottonseed 
      hydrolysate, as a medium additive, enhances Chinese hamster ovary (CHO) cell 
      culture performance. Metabolite profiling and tandem mass tag (TMT) proteomics 
      characterize its impact on CHO cells. Rewired nutrient utilization is observed 
      via glycolysis, amino acid, and polyamine metabolism. Hippo signaling pathway 
      impacts cell growth in the presence of cottonseed hydrolysate.
CI  - (c) 2023 Wiley-VCH GmbH.
FAU - Dhara, Venkata Gayatri
AU  - Dhara VG
AD  - Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 
      Baltimore, Maryland, USA.
FAU - Kumar, Swetha
AU  - Kumar S
AD  - Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 
      Baltimore, Maryland, USA.
FAU - DeVine, Lauren
AU  - DeVine L
AD  - Mass Spectrometry and Proteomics Facility, Johns Hopkins Medical Institute, 
      Baltimore, Maryland, USA.
FAU - Naik, Harnish Mukesh
AU  - Naik HM
AUID- ORCID: 0000-0002-8303-4219
AD  - Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 
      Baltimore, Maryland, USA.
FAU - Cole, Robert Norman
AU  - Cole RN
AD  - Mass Spectrometry and Proteomics Facility, Johns Hopkins Medical Institute, 
      Baltimore, Maryland, USA.
FAU - More, Abbie J
AU  - More AJ
AD  - FrieslandCampina Ingredients, Delhi, New York, USA.
FAU - Lau, Eduardo Catchon
AU  - Lau EC
AD  - FrieslandCampina Innovation, FrieslandCampina, Gelderland, Netherlands, Europe.
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
DEP - 20230411
PL  - Germany
TA  - Biotechnol J
JT  - Biotechnology journal
JID - 101265833
RN  - 0 (Cottonseed Oil)
RN  - 33X04XA5AT (Lactic Acid)
RN  - 8558G7RUTR (Pyruvic Acid)
RN  - 0 (Amino Acids)
RN  - 0 (Polyamines)
SB  - IM
MH  - Cricetinae
MH  - Animals
MH  - Cricetulus
MH  - CHO Cells
MH  - *Cottonseed Oil
MH  - *Proteomics
MH  - Batch Cell Culture Techniques
MH  - Lactic Acid/metabolism
MH  - Pyruvic Acid
MH  - Amino Acids/metabolism
MH  - Dietary Supplements
MH  - Polyamines
OTO - NOTNLM
OT  - CHO
OT  - bioprocessing
OT  - fold change analysis
OT  - hydrolysate
OT  - metabolism
OT  - proteomics
EDAT- 2023/03/10 06:00
MHDA- 2023/06/13 06:42
CRDT- 2023/03/09 08:53
PHST- 2023/02/16 00:00 [revised]
PHST- 2022/05/12 00:00 [received]
PHST- 2023/02/27 00:00 [accepted]
PHST- 2023/06/13 06:42 [medline]
PHST- 2023/03/10 06:00 [pubmed]
PHST- 2023/03/09 08:53 [entrez]
AID - 10.1002/biot.202200243 [doi]
PST - ppublish
SO  - Biotechnol J. 2023 Jun;18(6):e2200243. doi: 10.1002/biot.202200243. Epub 2023 Apr 
      11.