PMID- 30923861
OWN - NLM
STAT- MEDLINE
DCOM- 20190523
LR  - 20210107
IS  - 1618-2650 (Electronic)
IS  - 1618-2642 (Linking)
VI  - 411
IP  - 13
DP  - 2019 May
TI  - Simultaneous detection of nicotinamide adenine nucleotides and adenylate pool to 
      quantify redox and energy states in mAb-producing CHO cells by capillary 
      electrophoresis.
PG  - 2971-2979
LID - 10.1007/s00216-019-01747-9 [doi]
AB  - Chinese hamster ovary (CHO) cells are predominant in the production of 
      therapeutic proteins to treat various diseases. Characterization and 
      investigation of CHO cell metabolism in a quick and simple way could boost 
      process and cell line development. Therefore, a method to simultaneously detect 
      seven redox- and energy-related metabolites in CHO cells by capillary 
      electrophoresis has been developed. An on-line focusing technique was applied to 
      improve the peak shape and resolution by using a 50 mum x 44 cm uncoated fused 
      silica capillary. Key parameters and their interactions were investigated by 
      design of experiments (DoE) and optimized conditions were determined by 
      desirability function as follows: 24  degrees C, 95 mM, and pH 9.4 of BGE. The method was 
      validated to ensure sensitivity, linearity, and reproducibility. The limits of 
      detection (LODs) ranged from 0.050 to 0.688 mg/L for seven metabolites, and 
      correlation coefficients of linearity were all greater than 0.996. The relative 
      standard deviations (RSD) of migration time and peak area were smaller than 
      0.872% and 5.5%, respectively, except for NADPH, and the recoveries were between 
      97.5 and 101.2%. The method was successfully applied to analyze the extracts from 
      CHO cells under two different culture conditions. Graphical abstract.
FAU - Wang, Jiaqi
AU  - Wang J
AD  - State Key Laboratory of Bioreactor Engineering, East China University of Science 
      and Technology, Box 309, 130 Meilong Road, Shanghai, 200237, China.
FAU - Wang, Chen
AU  - Wang C
AD  - State Key Laboratory of Bioreactor Engineering, East China University of Science 
      and Technology, Box 309, 130 Meilong Road, Shanghai, 200237, China.
FAU - Fan, Li
AU  - Fan L
AD  - State Key Laboratory of Bioreactor Engineering, East China University of Science 
      and Technology, Box 309, 130 Meilong Road, Shanghai, 200237, China.
AD  - Shanghai BioEngine Sci-Tech Co. Ltd, Shanghai, 201203, China.
FAU - Zhao, Liang
AU  - Zhao L
AUID- ORCID: 0000-0001-8077-3617
AD  - State Key Laboratory of Bioreactor Engineering, East China University of Science 
      and Technology, Box 309, 130 Meilong Road, Shanghai, 200237, China. 
      zhaoliang@ecust.edu.cn.
FAU - Tan, Wen-Song
AU  - Tan WS
AD  - State Key Laboratory of Bioreactor Engineering, East China University of Science 
      and Technology, Box 309, 130 Meilong Road, Shanghai, 200237, China.
LA  - eng
PT  - Journal Article
DEP - 20190328
PL  - Germany
TA  - Anal Bioanal Chem
JT  - Analytical and bioanalytical chemistry
JID - 101134327
RN  - 0 (Antibodies, Monoclonal)
RN  - 0U46U6E8UK (NAD)
RN  - 415SHH325A (Adenosine Monophosphate)
RN  - 53-59-8 (NADP)
RN  - 61D2G4IYVH (Adenosine Diphosphate)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
SB  - IM
MH  - Adenosine Diphosphate/analysis
MH  - Adenosine Monophosphate/*analysis
MH  - Adenosine Triphosphate/analysis
MH  - Animals
MH  - Antibodies, Monoclonal/chemistry
MH  - CHO Cells
MH  - Cell Culture Techniques
MH  - Cricetulus
MH  - Electrophoresis, Capillary/*methods
MH  - Limit of Detection
MH  - NAD/*analysis
MH  - NADP/*analysis
MH  - Oxidation-Reduction
OTO - NOTNLM
OT  - CHO cells
OT  - Capillary electrophoresis
OT  - Design of experiments (DoE)
OT  - Nucleotides
OT  - On-line focusing
OT  - Redox
EDAT- 2019/03/30 06:00
MHDA- 2019/05/24 06:00
CRDT- 2019/03/30 06:00
PHST- 2018/12/25 00:00 [received]
PHST- 2019/03/01 00:00 [accepted]
PHST- 2019/02/15 00:00 [revised]
PHST- 2019/03/30 06:00 [pubmed]
PHST- 2019/05/24 06:00 [medline]
PHST- 2019/03/30 06:00 [entrez]
AID - 10.1007/s00216-019-01747-9 [pii]
AID - 10.1007/s00216-019-01747-9 [doi]
PST - ppublish
SO  - Anal Bioanal Chem. 2019 May;411(13):2971-2979. doi: 10.1007/s00216-019-01747-9. 
      Epub 2019 Mar 28.