A novel method for purifying bluetongue virus with high purity by co-immunoprecipitation with agarose protein A.
BACKGROUND
Bluetongue virus (BTV) is an icosahedral non-enveloped virus within the genus Orbivirus of Reoviridae and exists as 24 distinct serotypes. BTV can infect all ruminant species and causes severe sickness in sheep. Recently, it was reported that BTV can infect some human cancer cells selectively. Because of the important oncolysis of this virus, we developed a novel purifying method for large-scale production. The purifying logic is simple, which is picking out all the components unwanted and the left is what we want. The process can be summarized in 4 steps: centrifugation, pulling down cell debrises and soluble proteins by co-immunoprecipitation with agarose Protein A, dialysis and filtration sterilization after concentration.


RESULTS
The result of transmission electron microscope (TEM) observation showed that the sample of purified virus has a very clear background and the virions still kept intact. The result of 50% tissue culture infective dose (TCID(50)) assay showed that the bioactivity of purified virus is relatively high.


CONCLUSIONS
This method can purify BTV-10 with high quality and high biological activity on large-scale production. It also can be used for purifying other BTV serotypes.
Song
Zhen
Z
Lab. of Molecular Virus & Cancer, State Key Laboratory of Virology, Wuhan University School of Basic Medicine, Wuhan University, Wuhan 430071, China.
Dong
Changyuan
C
Wang
Lulu
L
Chen
Dong-E
DE
Bi
Guoming
G
Dai
Ming
M
Liu
Jun
J
D000818:Animals; D001819:Bluetongue / Q000821:virology*; D001820:Bluetongue virus / Q000302:isolation & purification* / Q000502:physiology / Q000648:ultrastructure; D002522:Chlorocebus aethiops; D047468:Immunoprecipitation / Q000379:methods*; D011485:Protein Binding; D012685:Sepharose / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry*; D014709:Vero Cells; D014773:Virology / Q000379:methods*
D013205:Staphylococcal Protein A; D012685:Sepharose