PMID- 17913250
OWN - NLM
STAT- MEDLINE
DA  - 20071218
DCOM- 20080318
LR  - 20101118
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 147
IP  - 1
DP  - 2008 Jan
TI  - Purification of the major envelop protein GP5 of porcine reproductive and
      respiratory syndrome virus (PRRSV) from native virions.
PG  - 127-35
AB  - Porcine reproductive and respiratory syndrome virus (PRRSV) is the cause of an
      economically important swine disease that has been devastating the global swine
      industry since the early 1990s. The current PRRSV vaccines are not very effective
      largely due to heterogeneic nature of the virus. The major envelope protein, GP5,
      exposes outside the virion, induces neutralizing antibodies, and thus is a
      primary target for developing a subunit vaccine. In this study, we report a
      process for purification of GP5 protein from native virions of PRRSV propagated
      in MARC-145 cells. PRRSV virions were first purified and concentrated through
      sucrose cushion ultracentrifugation. GP5 protein was subsequently solubilized
      with Triton X-100 detergent for further processing. Cation exchange
      chromatography (CEX) was utilized for partial fractionation of GP5, although the 
      viral nucleocapsid protein (N) was a major impurity in CEX elution fractions.
      During a second chromatographic step, hydrophobic interaction chromatography
      (HIC) further purified GP5 protein by means of a two-stage elution scheme. Pure
      GP5 protein was eluted from the HIC resin in the second HIC elution stage by
      Triton X-100 displacement; however the protein is present as a
      homodimeric/tetrameric aggregate. This process may be useful in PRRSV subunit
      vaccine development.
AD  - Department of Biological Systems Engineering, Virginia Polytechnic Institute and 
      State University, 200 Seitz Hall, Blacksburg, VA 24061, USA.
FAU - Matanin, Brad M
AU  - Matanin BM
FAU - Huang, Yaowei
AU  - Huang Y
FAU - Meng, X J
AU  - Meng XJ
FAU - Zhang, Chenming
AU  - Zhang C
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20071025
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (Viral Vaccines)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Chromatography/*methods
MH  - Chromatography, Ion Exchange/methods
MH  - Hydrophobic and Hydrophilic Interactions
MH  - Porcine respiratory and reproductive syndrome
      virus/*chemistry/immunology/isolation & purification/physiology
MH  - Viral Envelope Proteins/immunology/*isolation & purification
MH  - Viral Vaccines
MH  - Virion/chemistry
MH  - Virus Cultivation
EDAT- 2007/10/05 09:00
MHDA- 2008/03/19 09:00
CRDT- 2007/10/05 09:00
PHST- 2007/06/12 [received]
PHST- 2007/08/17 [revised]
PHST- 2007/08/22 [accepted]
PHST- 2007/10/25 [aheadofprint]
AID - S0166-0934(07)00332-1 [pii]
AID - 10.1016/j.jviromet.2007.08.018 [doi]
PST - ppublish
SO  - J Virol Methods. 2008 Jan;147(1):127-35. Epub 2007 Oct 25.