PMID- 20371065
OWN - NLM
STAT- MEDLINE
DCOM- 20100729
LR  - 20151119
IS  - 1873-3778 (Electronic)
IS  - 0021-9673 (Linking)
VI  - 1217
IP  - 21
DP  - 2010 May 21
TI  - Purification of porcine reproductive and respiratory syndrome virus from cell 
      culture using ultrafiltration and heparin affinity chromatography.
PG  - 3489-93
LID - 10.1016/j.chroma.2010.03.023 [doi]
AB  - Porcine reproductive and respiratory syndrome (PRRS) virus is the causative agent 
      of the most significant infectious disease currently affecting the swine industry 
      worldwide. Density gradient ultracentrifugation remains the most commonly used 
      method for porcine reproductive and respiratory syndrome virus (PRRSV) 
      purification. However, this technique has notable drawbacks including long 
      processing time and limited processing volume in each run. To overcome these 
      limitations, a scalable process was developed. PRRSV propagated in MARC-145 was 
      released by three freeze/thaw cycles. After a low speed centrifugation step, the 
      virus particles in the supernatant were concentrated twice by an ultrafiltration 
      step. The ultrafiltration step concentrated the virions effectively with no 
      detectable loss while some cultural/cellular proteins were removed. The virions 
      in the ultrafiltration retentate were then applied to a heparin affinity column 
      on a fast performance liquid chromatography unit. The combined ultrafiltration 
      and heparin affinity chromatography process removed more than 96% of cellular and 
      medium proteins. During a stepwise elution strategy, the viral particles were 
      eluted at two separate peaks recovering 27.5% and 25.4% of viral particles loaded 
      onto the column with a purity of 194 and 3917 particles/microg protein, 
      respectively.
CI  - 2010 Elsevier B.V. All rights reserved.
FAU - Hu, Jianzhong
AU  - Hu J
AD  - Department of Biological Systems Engineering, Virginia Polytechnic Institute and 
      State University, 200 Seitz Hall, Blacksburg, VA 24061, USA.
FAU - Ni, Yanyan
AU  - Ni Y
FAU - Dryman, Barbara A
AU  - Dryman BA
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 - 20100319
PL  - Netherlands
TA  - J Chromatogr A
JT  - Journal of chromatography. A
JID - 9318488
RN  - 023C2WHX2V (Tromethamine)
RN  - 451W47IQ8X (Sodium Chloride)
RN  - 9005-49-6 (Heparin)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Cell Line
MH  - Chromatography, Affinity/*methods
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Freezing
MH  - Heparin/metabolism
MH  - Porcine Reproductive and Respiratory Syndrome/virology
MH  - Porcine respiratory and reproductive syndrome virus/*isolation & purification
MH  - Sodium Chloride
MH  - Swine
MH  - Tromethamine
MH  - Ultrafiltration/*methods
MH  - Virion/*isolation & purification/metabolism
MH  - Virus Cultivation/*methods
EDAT- 2010/04/08 06:00
MHDA- 2010/07/30 06:00
CRDT- 2010/04/08 06:00
PHST- 2009/11/06 00:00 [received]
PHST- 2010/03/10 00:00 [revised]
PHST- 2010/03/15 00:00 [accepted]
PHST- 2010/04/08 06:00 [entrez]
PHST- 2010/04/08 06:00 [pubmed]
PHST- 2010/07/30 06:00 [medline]
AID - S0021-9673(10)00387-0 [pii]
AID - 10.1016/j.chroma.2010.03.023 [doi]
PST - ppublish
SO  - J Chromatogr A. 2010 May 21;1217(21):3489-93. doi: 10.1016/j.chroma.2010.03.023. 
      Epub 2010 Mar 19.