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Study on the purification of OPV produced on vero cells with Q-Sepharose F. F.
Authors:Liao Guo-yang  Jiang Shu-de  Zheng Cong-yi  Wang Ming-xiu  Li Ping-zhong
Institution:(1) Institute of Medical Biology, Chinese Academy of Medical Science & Peking Union of Medical College, 650107 Kunming, China;(2) College of Life Sciences, Wuhan University, 430072 Wuhan, China
Abstract:Q-Sepharose Fast Flow (Q-Sepharose F. F.) was adopted to purify the suspensions of type I, II, III oral poliovaccine (OPV), which is prepared from Vero cells. After clarification and concentration by ultrafiltration, the recovery of virus infected titre may attain above 85%. Using column chromatography on Q-Sepharose F. F. to purify the concentrated virus suspensions, the purified viruses attain 100% recovery of virus infectivity. Dot membrane hybrization was used to detect DNA with the probe of Vero cell genome DNA which was labeled with α-P32 dATP, the contents of residual substrate DNA was less than 100 Pg/dose. The process of downstream had no significant influence on some biological characters of purified viruses, such as virus morphology, tumorigenicity, rct/40 character and capsid protein. This downstream reseach indicates that Q-Sepharose F. F. is an ideal chromatography material for purifying OPV prepared from Vero cells. Biography: LIAO Guo-yang (1962-), Ph D Candidate.
Keywords:OPV  vero cells  substrate DNA  purification  Q-Sepharose F  F  
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