tailieunhanh - Báo cáo y học: "A novel method for purifying bluetongue virus with high purity by co-immunoprecipitation with agarose protein A"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: A novel method for purifying bluetongue virus with high purity by co-immunoprecipitation with agarose protein A | Zhen et al. Virology Journal 2010 7 126 http content 7 1 126 VIROLOGY JOURNAL METHODOLOGY Open Access A novel method for purifying bluetongue virus with high purity by co-immunoprecipitation with agarose protein A Song Zhen 1 Dong Changyuan 1 Wang Lulu1 Chen Dong-E2 Bi Guoming1 Dai Ming1 and Liu Jun1 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 TCID50 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. Background Bluetongue virus BTV is an icosahedral non-enveloped virus within the genus Orbivirus of Reoviridae 1 and exists as 24 distinct serotypes 2 . BTV can infect ruminant species mainly through the biting Culicoides species 3 . Particularly it usually causes severe bluetongue diseases BT only in sheep and some species of deer. Over the past century BTV has never shown infectivity on Homosapiens and no normal human being cells have been successfully