PMID- 23885665
OWN - NLM
STAT- MEDLINE
DCOM- 20140213
LR  - 20130802
IS  - 1873-3778 (Electronic)
IS  - 0021-9673 (Linking)
VI  - 1304
DP  - 2013 Aug 23
TI  - Purification and characterization of recombinant envelope protein GP5 of porcine 
      reproductive and respiratory syndrome virus from E. coli.
PG  - 133-7
LID - S0021-9673(13)01022-4 [pii]
LID - 10.1016/j.chroma.2013.06.079 [doi]
AB  - The major envelope protein, GP5, in porcine reproductive and respiratory syndrome 
      virus (PRRSV) plays critical roles in the assembly, invasion and immune response 
      of PRRSV particle, and is one of the mostly studied candidates in the development 
      of recombinant vaccines. In this research, a purification process including 
      immobilized metal affinity chromatography (IMAC) and hydrophobic interaction 
      chromatography (HIC) was developed to prepare recombinant envelope protein GP5 
      with His-tag from an E. coli strain transformed with pGEM-ORF5. The result of 
      cell culture indicated that His-tagged GP5 protein was expressed mainly in 
      soluble form. After cell disruption, His-tagged GP5 protein was successfully 
      purified by Ni(2+)-chelating Sepharose Fast Flow with a yield and purity of 80.5% 
      and 48%, respectively. Recombinant GP5 protein was further purified by HIC to 
      achieve a purity of 95%. Moreover, the purified rGP5 is shown in monomeric form 
      contrasting the dimeric or tetrameric form when purified by a CEX-HIC process 
      directly from PRRSV virions as reported in a previous study.
CI  - Copyright (c) 2013 Elsevier B.V. All rights reserved.
FAU - Liu, Chong
AU  - Liu C
AD  - Institute of Chemical Engineering, School of Chemical Engineering and Technology, 
      Tianjin University, Tianjin 300072, PR China.
FAU - Liu, Chun-Jiang
AU  - Liu CJ
FAU - Yuan, Xi-Gang
AU  - Yuan XG
FAU - Zhang, Chenming
AU  - Zhang C
LA  - eng
PT  - Journal Article
DEP - 20130704
PL  - Netherlands
TA  - J Chromatogr A
JT  - Journal of chromatography. A
JID - 9318488
RN  - 0 (Metals)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Vaccines, Synthetic)
RN  - 0 (Viral Envelope Proteins)
SB  - IM
MH  - Animals
MH  - Chromatography, Affinity/*methods
MH  - Cloning, Molecular
MH  - Escherichia coli/genetics
MH  - Hydrophobic and Hydrophilic Interactions
MH  - Metals/chemistry
MH  - Porcine Reproductive and Respiratory Syndrome/*virology
MH  - Porcine respiratory and reproductive syndrome virus/*chemistry/genetics
MH  - Recombinant Proteins/chemistry/genetics/*isolation & purification
MH  - Swine
MH  - Vaccines, Synthetic/chemistry/genetics/isolation & purification
MH  - Viral Envelope Proteins/chemistry/genetics/*isolation & purification
OTO - NOTNLM
OT  - Envelope protein GP5
OT  - Histidine-tag
OT  - Hydrophobic interaction chromatography
OT  - Immobilized metal affinity chromatography
OT  - Porcine reproductive and respiratory syndrome virus
EDAT- 2013/07/28 06:00
MHDA- 2014/02/14 06:00
CRDT- 2013/07/27 06:00
PHST- 2013/04/19 00:00 [received]
PHST- 2013/06/27 00:00 [revised]
PHST- 2013/06/30 00:00 [accepted]
PHST- 2013/07/27 06:00 [entrez]
PHST- 2013/07/28 06:00 [pubmed]
PHST- 2014/02/14 06:00 [medline]
AID - S0021-9673(13)01022-4 [pii]
AID - 10.1016/j.chroma.2013.06.079 [doi]
PST - ppublish
SO  - J Chromatogr A. 2013 Aug 23;1304:133-7. doi: 10.1016/j.chroma.2013.06.079. Epub 
      2013 Jul 4.