摘要
地鼠肾细胞狂犬病疫苗原液经１００ ｋＤ 膜浓缩 ３０ 倍，分别选用（１）ＤＥＡＥ Ｓｅｐｈａｒｏｓｅ ＣＬ－６Ｂ离子交换层析法；（２）Ｓｅｐｈａｃｒｙ１ Ｓ－２００ ＨＲ 分子筛选层析法；（３）二次蔗糖等密度区带离心法对其进行纯化。用此三种方法各试制３ 批精制疫苗，结果表明，经ＤＥＡＥ Ｓｅｐｈａｒｏｓｅ ＣＬ－６Ｂ离子交换层析纯化后疫苗总蛋白含量减少９９％ 以上，抗原比活性提高１５９ 倍，抗原回收率达５０％ ，纯化疫苗以ＮＩＨ 法效力测定平均为５．４ ＩＵ／２ｍ ｌ；经Ｓｅｐｈａｃｒｙ１ Ｓ－２００ＨＲ 分子筛层析纯化后疫苗总蛋白含量减少 ９８％ 以上，抗原比活性提高４１ 倍，抗原回收率达６３％ ，纯化疫苗效力平均为６．２５ ＩＵ／２ｍ ｌ；经一次蔗糖等密度区带离心法纯化后疫苗总蛋白含量减少９８％ 以上，抗原比活性提高３２１ 倍，抗原回收率达４３％ ，纯化疫苗效力平均为６．１８ ＩＵ／２ｍ ｌ，三种纯化疫苗均符合Ｗ ＨＯ 规程要求。
The rabies vaccine,concentrated 30 times by 100 kD membrane,were purified by(1)DEAE Sepharose CL 6B chromatography,(2)Sephacryl S 200HR chromatography,and(3)twice sucrose density gradient centrifugation methods.Three lots vaccines were prepared separately by above methods.The presults showed that,with DEAE Sepharose CL 6B chromatography,the unneeded protein decreased more than 99%,the antigen recovery is about 50%,and the NIH potency reaches to 5.4 IU/2ml;with Sephacryl S 200 HR chromatography,the protein decreased more than 98%,the antigen recovery is about 63%,and the NIH potency reaches to 6.25 IU/2ml.While the centrifugation method gave a protein decrease of 98%,antigen recovery of 43% and NIH potency of 6.18 IU/2ml.All of the purified vaccines can meet the requirements WHO.
出处
《微生物学免疫学进展》
1999年第4期37-41,共5页
Progress In Microbiology and Immunology
关键词
精制
狂犬病疫苗
纯化
比较
Purified rabies vaccine Purification Comparison
