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Zhou L.-B.,Liaoning Chengda Biological Technique Co. | Yin J.-W.,Liaoning Chengda Biological Technique Co. | Zhao X.,Liaoning Chengda Biological Technique Co. | Wu X.-T.,Liaoning Chengda Biological Technique Co. | And 6 more authors.
Chinese Journal of Biologicals | Year: 2012

Objective: To culture Vero cells and Japanese encephalitis virus (JEV) and produce inactivated JE vaccine in a large scale by using 7.5 L bioreactor. Methods: A 7.5 L bioreactor was loaded with microcarriers at densities of 5, 10, 15, 20 and 25 g/L respectively, on which Vero cells were cultured by continuous perfusion and inoculated with JEV P3 strain at MOIs of 0.005 00, 0.002 50, 0.001 60, 0.001 25 and 0.001 00 respectively. The virus liquid was harvested, concentrated, inactivated and purified to prepare the final bulk of JE vaccine, and the procedure was verified by vaccine inactivation and purification tests. The final bulk qualified in control tests were distributed to obtain final product which was subjected to overall control tests according to the requirements in monograph of Japanese Encephalitis Vaccine (Vero cell), Inactivated, Freeze-dried in Chinese Pharmacopoeia (Volume III, 2010 edition). Results: When the densities of microcarriers were 20 and 25 g/L, the density of Vero cells reached 1.2 × 107 cells/ml, while the mean virus titer was 8.33-8.52 IgLD50/ml. However, when the MOI of virus was 0.001 25, the virus titer reached 9.02 IgLD50/ml at most. The virus bulk concentrated by ultrafiltration was effectively inactivated after treatment with β-propionolactone at a dilution of 1 : 4 000 for 72 h. Ninety percent of foreign protein was removed after purification. All the quality indexes of six batches of vaccine prepared by using 7.5 L bioreactor met the relevant national requirements. Conclusion: Inactivated JE vaccine was produced in a large scale by inoculation of JEV P3 strain into Vero cells cultured in 7.5 L bioreactor by continuous perfusion.

Yang W.-Y.,Liaoning Chengda Biological Technique Co. | Xiu X.-L.,Liaoning Chengda Biological Technique Co. | Wang Y.,Liaoning Chengda Biological Technique Co. | Zhang J.,Liaoning Chengda Biological Technique Co. | And 3 more authors.
Chinese Journal of Biologicals | Year: 2016

Objective To prepare adsorbed Proteus miral>ilLi-Staphylococcus aureus-Pseudomonas aeruginosa (PSP) combined vaccine and investigate its protective effect on mice. Methods I'roteus mirabilis membrane antigen (PMA), Staphylococcus aureus cytoplasmic antigen (SCA), Staphylococcus aureus toxoid antigen (SEA) and Pseudomonas aeruginosa toxoid antigen (PEA) were prepared by different procedures, and formulated with aluminum hydroxide at a certain ratio to obtain adsorbed PSP combined vaccine, of which the protective effect was measured by immunization of mice by intraperitoneal route. Results The protective rates of hexavalent PMA to were 50% ∼ 100% to six I'roteus miraliilis vaccine strains, while those of adsorbed I'SP combined vaccine were 80% to I'roteus mirabilis, 100% to P. aeruginosa PA 101 strain, 90% and 60% to S. aureus SA79 and CMCC26002 strains respectively, and 70% to a-Toxin of CMCC26002 strain. Of the other strains of the same group to which the protective rate of PSP were more than 70%, 61. 11% were P. miral>ilis, 50% were S. aureus, and 92. 86% were P. aeruginosa. Conclusion Adsorbed PSP combined vaccine showed protective effect on the vaccine strains, while showed also protective effect on other strains of the same group, which provided a theoretical basis and technical support for further clinical trials.

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