PMID- 24039059
OWN - NLM
STAT- MEDLINE
DCOM- 20141016
LR  - 20211021
IS  - 1860-7314 (Electronic)
IS  - 1860-6768 (Print)
IS  - 1860-6768 (Linking)
VI  - 9
IP  - 1
DP  - 2014 Jan
TI  - Recovery of Chinese hamster ovary host cell proteins for proteomic analysis.
PG  - 87-99
LID - 10.1002/biot.201300190 [doi]
AB  - Identification and characterization of Chinese hamster ovary (CHO) host cell 
      protein (HCP) impurities by proteomic techniques can aid bioprocess design and 
      lead to more efficient development and improved biopharmaceutical manufacturing 
      operations. Recovery of extracellular CHO HCP for proteomic analysis is 
      particularly challenging due to the relatively low protein concentration and 
      complex composition of media. In this article, we report the development of 
      optimized protocols that improve proteome capture for CHO HCP. Eleven 
      precipitation protocols were screened for protein recovery and optimized for a 
      subset of precipitants by a design of experiments (DOE) approach. Because total 
      protein recovery does not fully replicate a proteomics experiment, or detect 
      non-protein agents that may interfere with proteomic methods, a subset of 
      precipitation conditions were compared by two-dimensional electrophoresis and 
      liquid chromatography coupled with mass spectrometry, with optimized recovery 
      shown to differ between the two proteomic methods. This work demonstrates broadly 
      applicable methods that can be applied as initial steps to optimize sample 
      preparation of any sample type for proteomic analysis, and presents optimized 
      precipitation protocols for extracellular CHO HCP recovery, which can vary 
      appreciably between gel-based and shotgun proteomic methods.
CI  - Copyright (c) 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
FAU - Valente, Kristin N
AU  - Valente KN
AD  - Department of Chemical and Biomolecular Engineering, University of Delaware, 
      Newark, DE, USA; Delaware Biotechnology Institute, University of Delaware, 
      Newark, DE, USA.
FAU - Schaefer, Amy K
AU  - Schaefer AK
FAU - Kempton, Hannah R
AU  - Kempton HR
FAU - Lenhoff, Abraham M
AU  - Lenhoff AM
FAU - Lee, Kelvin H
AU  - Lee KH
LA  - eng
GR  - T32 GM008550/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131210
PL  - Germany
TA  - Biotechnol J
JT  - Biotechnology journal
JID - 101265833
RN  - 0 (Proteome)
RN  - 0 (Solvents)
SB  - IM
MH  - Animals
MH  - CHO Cells/*chemistry
MH  - Cricetinae
MH  - Cricetulus
MH  - Electrophoresis, Gel, Two-Dimensional/methods
MH  - High-Throughput Screening Assays/*methods
MH  - Proteome/*chemistry/*isolation & purification
MH  - Proteomics/*methods
MH  - Research Design
MH  - Solvents
PMC - PMC5885764
MID - NIHMS951562
OTO - NOTNLM
OT  - Chinese hamster ovary cells
OT  - Design of experiments optimization
OT  - Extracellular host cell proteins
OT  - Proteomics
OT  - Sample preparation
COIS- Conflict-of-interest statement The authors declare no commercial or financial 
      conflict of interest.
EDAT- 2013/09/17 06:00
MHDA- 2014/10/17 06:00
PMCR- 2018/04/05
CRDT- 2013/09/17 06:00
PHST- 2013/04/30 00:00 [received]
PHST- 2013/07/03 00:00 [revised]
PHST- 2013/08/21 00:00 [accepted]
PHST- 2013/09/17 06:00 [entrez]
PHST- 2013/09/17 06:00 [pubmed]
PHST- 2014/10/17 06:00 [medline]
PHST- 2018/04/05 00:00 [pmc-release]
AID - 10.1002/biot.201300190 [doi]
PST - ppublish
SO  - Biotechnol J. 2014 Jan;9(1):87-99. doi: 10.1002/biot.201300190. Epub 2013 Dec 10.