title: A novel method for purifying bluetongue virus with high purity by co-immunoprecipitation with agarose protein A.
abstract: 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.
authors.affiliation: Lab. of Molecular Virus & Cancer, State Key Laboratory of Virology, Wuhan University School of Basic Medicine, Wuhan University, Wuhan 430071, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Song
authors.forename: Changyuan
authors.initials: C
authors.lastname: Dong
authors.forename: Lulu
authors.initials: L
authors.lastname: Wang
authors.forename: Dong-E
authors.initials: DE
authors.lastname: Chen
authors.forename: Guoming
authors.initials: G
authors.lastname: Bi
authors.forename: Ming
authors.initials: M
authors.lastname: Dai
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
mesh_terms: 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*
chemical_list: D013205:Staphylococcal Protein A; D012685:Sepharose