PMID- 23217304
OWN - NLM
STAT- MEDLINE
DCOM- 20130429
LR  - 20131121
IS  - 1873-376X (Electronic)
IS  - 1570-0232 (Linking)
VI  - 911
DP  - 2012 Dec 12
TI  - Expression and purification of a chimeric protein consisting of the ectodomains 
      of M and GP5 proteins of porcine reproductive and respiratory syndrome virus 
      (PRRSV).
PG  - 43-8
LID - S1570-0232(12)00598-3 [pii]
LID - 10.1016/j.jchromb.2012.10.014 [doi]
AB  - Porcine reproductive and respiratory syndrome (PRRS) is the most economically 
      important infectious disease currently affecting the swine industry worldwide. In 
      the US alone, it causes economic losses of more than 560 million dollars every 
      year. Although killed-virus and modified-live PRRS vaccines are commercially 
      available, the unsatisfactory efficacy and safety of current vaccines drives the 
      impetus of developing novel PRRSV vaccines. To fulfill this purpose, we designed 
      a chimeric protein consisting of the ectodomains of viral GP5 and M protein, the 
      two most widely studied subunit vaccine targets, and expressed it in E. coli. An 
      optimized purification/refolding process composed of immobilized metal ion 
      affinity chromatography, dialysis refolding and anion exchange chromatography was 
      developed to purify the chimeric protein from the inclusion bodies. This process 
      could recover approximately 12mgprotein/l E. coli broth with near 100% purity and 
      very low endotoxin level. In addition, the purified protein is antigenic, can 
      bind to a cellular receptor for the virus (heparan sulfate), and can block virus 
      infection of susceptible cells. Therefore, the chimeric protein is a promising 
      subunit vaccine candidate against PRRSV.
CI  - Copyright (c) 2012 Elsevier B.V. All rights reserved.
FAU - Hu, Jianzhong
AU  - Hu J
AD  - Department of Biological Systems Engineering, Virginia Tech, 200 Seitz Hall, 
      Blacksburg, VA 24061, USA.
FAU - Ni, Yanyan
AU  - Ni Y
FAU - Meng, X J
AU  - Meng XJ
FAU - Zhang, Chenming
AU  - Zhang C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20121017
PL  - Netherlands
TA  - J Chromatogr B Analyt Technol Biomed Life Sci
JT  - Journal of chromatography. B, Analytical technologies in the biomedical and life 
      sciences
JID - 101139554
RN  - 0 (His-His-His-His-His-His)
RN  - 0 (M protein, porcine reproductive and respiratory syndrome virus)
RN  - 0 (Oligopeptides)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (Viral Matrix Proteins)
RN  - 0 (glycoprotein 5, PRRSV)
RN  - 4QD397987E (Histidine)
RN  - 9050-30-0 (Heparitin Sulfate)
SB  - IM
MH  - Blotting, Western
MH  - Chromatography, Ion Exchange/methods
MH  - Escherichia coli/chemistry/genetics/metabolism
MH  - Heparitin Sulfate/chemistry
MH  - Histidine/genetics/metabolism
MH  - Oligopeptides/genetics/metabolism
MH  - Protein Refolding
MH  - Protein Structure, Tertiary
MH  - Recombinant Fusion Proteins/*biosynthesis/genetics/*isolation & purification
MH  - Viral Envelope Proteins/*isolation & purification/metabolism
MH  - Viral Matrix Proteins/*biosynthesis/*isolation & purification/metabolism
EDAT- 2012/12/12 06:00
MHDA- 2013/04/30 06:00
CRDT- 2012/12/11 06:00
PHST- 2012/08/14 00:00 [received]
PHST- 2012/09/21 00:00 [revised]
PHST- 2012/10/10 00:00 [accepted]
PHST- 2012/12/11 06:00 [entrez]
PHST- 2012/12/12 06:00 [pubmed]
PHST- 2013/04/30 06:00 [medline]
AID - S1570-0232(12)00598-3 [pii]
AID - 10.1016/j.jchromb.2012.10.014 [doi]
PST - ppublish
SO  - J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Dec 12;911:43-8. doi: 
      10.1016/j.jchromb.2012.10.014. Epub 2012 Oct 17.