PMID- 25215178
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20140912
LR  - 20220330
IS  - 2008-2835 (Print)
IS  - 2008-4625 (Electronic)
IS  - 2008-2835 (Linking)
VI  - 6
IP  - 3
DP  - 2014 Jul
TI  - Effect of Peptone Feeding on Transient Gene Expression Process in CHO DG44.
PG  - 147-55
AB  - BACKGROUND: Transient Gene Expression (TGE) gained popularity over the last 
      decade as a rapid method for the production of milligram to gram quantities of 
      recombinant proteins for preclinical studies in biophama industry. Thereby, the 
      optimization of the TGE technique for Chinese hamster ovary (CHO) as the dominant 
      host for the production of biotherapeutics is of great interest to reach the 
      values for Human Embryo Kidney-293 (HEK-293) cells in terms of transfection 
      efficiencies and production titers. TGE efficiencies are cell line and vector 
      dependant. METHODS: In transfection efficiency optimization experiments, 
      different starting cell densities, different amounts of plasmid DNA and PEI 
      transfection reagent were investigated to achieve the best conditions leading to 
      maximum transfection efficiencies. Furthermore, in order to investigate the 
      effect of peptone feeding on transfection efficiency, three different sources of 
      peptones with the greatest effect in the CD DG44 basal media were selected; 
      Casein Tryptone N1, Soy petone A2SC and Soy peptone E110. RESULTS: The 
      transfection strategy performed here was able to make an outstanding increase in 
      transfection efficiency of CHO DG44 cell line transfected with 
      pTracer-SV40-mutated t-PA plasmid from 3.6% in our starting non-optimized 
      condition to 66.93% in finally optimized situation. Moreover, peptone feeding 
      strategy used here was successful to increase volumetric productivities up to 
      37%. In addition, the amounts of both PEI and plasmid DNA were reduced up to 66% 
      and 25%, respectively compared to our previous protocol. CONCLUSION: Here we 
      described an optimization process for TGE in suspension-adapted CHO cells based 
      on Polyethylenimine (PEI)/DNA concentration, DNA: PEI ratio, starting cell 
      densities and peptone feeding strategy.
FAU - Davami, Fatemeh
AU  - Davami F
AD  - Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
FAU - Eghbalpour, Farnaz
AU  - Eghbalpour F
AD  - Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran ; Islamic 
      Azad University of Arak, Arak, Iran.
FAU - Barkhordari, Farzaneh
AU  - Barkhordari F
AD  - Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
FAU - Mahboudi, Fereidoun
AU  - Mahboudi F
AD  - Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
LA  - eng
PT  - Journal Article
PL  - Iran
TA  - Avicenna J Med Biotechnol
JT  - Avicenna journal of medical biotechnology
JID - 101511065
PMC - PMC4147101
OTO - NOTNLM
OT  - Chinese hamster ovary cells
OT  - Peptones
OT  - Polyethylenimine
OT  - Protein production
OT  - Transient gene expression
EDAT- 2014/09/13 06:00
MHDA- 2014/09/13 06:01
PMCR- 2014/07/01
CRDT- 2014/09/13 06:00
PHST- 2014/01/12 00:00 [received]
PHST- 2014/03/08 00:00 [accepted]
PHST- 2014/09/13 06:00 [entrez]
PHST- 2014/09/13 06:00 [pubmed]
PHST- 2014/09/13 06:01 [medline]
PHST- 2014/07/01 00:00 [pmc-release]
AID - AJMB-6-147 [pii]
PST - ppublish
SO  - Avicenna J Med Biotechnol. 2014 Jul;6(3):147-55.