柱层析法纯化人用地鼠肾细胞狂犬病毒灭活疫苗的研究

目的改进和提高现行人用地鼠肾细胞狂犬病灭活疫苗的纯度和保护效价。方法　将原代地鼠肾细胞 (PHKC)培养并收获的aG株狂犬病毒原液超滤浓缩 40～ 5 0倍 ,再经Sepharose 6FastFlow分子筛柱层析 ,收集完整的病毒颗粒 ,除菌、灭活、添加Al (OH) 3 佐剂制成纯化地鼠肾细胞狂犬病毒疫苗。结果　浓缩狂犬病毒经过柱层析纯化 ,抗原比活性较纯化前平均提高 2 9 5 4倍 ,纯化效果平均达到 96 1% ,疫苗效力平均达到 2 98IU/ml,其余检测结果均符合现行有关规程。结论　凝胶分子筛柱层析技术 ,是一种快捷而经济的改进现行狂犬疫苗质量的有效方法

STUDY ON HUMAN RABIES VACCINE PURIFIED BY COLUMN CHROMATOGRAPHY

Aim To improve the purity and protection potency of the current primary hamster kidney cell(PHKC) rabies vaccine of human use.Methods The suspension of rabies virus of aG strain obtained from PHKC was concentrated 40～50 times by ultrafiltration with a cut-off of 300,000 and purified by column chromatography on Sepharose 6 Fast Flow. The purified rabies virus was collected, sterilized and inactivated, then added in aluminium hydroxide to formulate the purified PHKC rabies vaccine.Results After concentration and column chromatography on Seepharose 6 Fast Flow, the average specific activity of the virus was improved by 29.54 times, the average purification efficiency reached to 96 1% and NIH potency about 2 98IU/ml.Its potency recovery after purification reached to 57 6%. All the other test results met the demands of WHO and our regulation. Conclusion The purity and potency of the current rabies vaccine was improved rapidly and economically by column chromatography on Sepharose 6 FF.