People With Somatic Cell Rabies Vaccine Purification Technology Research

The significance of this esearch:rabies epidemic in China has been on an upward trend,the number of death is the second in the world and first in 37 kinds of notifiable infectious diseases in our country.At present,the rabies vaccine is the only effective drug of prevention and treatment of Zoonosis rabies?That vaccine were cultured by animal cells,which has potential risk of residual tumor induced DNA.With the human diploid cells to culture virus,preparation of rabies vaccine is by far the best rabies vaccine,known as the gold standard for vaccine.Normal cells human diploid cells have not t residual cancer risk of other animal cells,and its immunogenicity is very high,so it is only used in America,Canada,most European countries and some Asian countriesTo explore the research of the production process quality control and assurance test system of human diploid cell rabies vaccine,on the basis of the previous work,furtherly improve the safety and efficacy of the vaccine.Experiment method:Tt is played an important role that the production process of ensuring the quality of the vaccine,especially the quantitative indicators in the production process is more important.Design of experiment is tested Cell sensitivity to rabies virus,respectively.The CTN-1V strain was adapted in human embryo lung cell line Walvax-2,preparation of liquid culture of rabies virus cells by the method of using Walvax-2 cell line cell factori-es,ultrafiltrated concentration of rabies virus by compared to 300KD membrane,300KD,750KD hollow fiber,selected the appropriate concentration,the purification after concentrated liquid by Capto 700 column chromatography,Sepharose 4FF,the gel ion exchange DEAE Sepharose FF.Select the appropriate conditions comparing each column chromatography purification result,and the preliminary exploration of the beta propiolac-tone in column chromatography before and after inactivation due to the degeneration of human albumin and sensitization has carried.results:the the virus titer of P1 generation was 3.01ogLD50/ml and above P157.01ogLD50/ml in the 15 generation.Gained the rabies vaccine of being sensitive to human diploid cell.Preparaed the 7.51ogLD50/ml rabies virus titer,Results also shows better effect with 750KD hollow fiber by Ultrafiltration virus stock solution effectively removed impurities and high recovery rate of protein through Sepharose 4FF,Capto Core gel composite 700 column chromatography excellent quality rabies vaccine is obtain in mean of purified and beta propiolactone inactivation And preparaed the better rabies vaccine stock solution of a human diploid cell in the various indicators...