犬细小病毒层析纯化方法的建立OACSTPCD

The purity of the virus is the key basis for the preparation of antibodies and the improvement of vaccine immunity.In this paper,methods for purifying canine parvovirus(CPV)were established with Purose shell V15 ion-exchange chromatography and Sepharose 4FF molecular sieve chromatography.CPV was clarified by centrifugation and was concentrated with tangential flow filtration.Then,virus purifica-tion was performed on AKTA purifier 100 combined with Purose shell V15 or Sepharose 4FF,and was next concentrated by ultrafiltration spin columns.Finally,the virus content was analyzed with hemagglutination test,total protein removal rate was determined by BCA assay,and the purified virus was determined by SDS-PAGE,Western blot test and electron microscope observation.The results showed that the parvovirus purified with the two methods was typical in shape,and the diameter of the virus particle was about 20 nm.The total protein removal rate was over 97%.The virus purified by Purose shell V15 showed no disturbing bands of cellular components in the SDS-PAGE and Western blot ex-periments,while the virus purified by Sepharose 4FF had a few bovine serum albumin bands.The experiments showed that both the methods can be used for the purification of CPV,and the virus purified by Purose shell V15 was of higher purity.