Study on the purification of OPV produced on vero cells with Q-Sepharose F. F. |
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Authors: | Liao Guo-yang Jiang Shu-de Zheng Cong-yi Wang Ming-xiu Li Ping-zhong |
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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 |
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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. |
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Keywords: | OPV vero cells substrate DNA purification Q-Sepharose F F |
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