摘要
用乙型肝炎病毒(HBV)DNA转染工程细胞株MT-5,收获培养上清液,经超过滤浓缩、PEG沉淀和3次超速离心,得到纯化的HBsAg。经SDS-PAGE银染色后,纯化的HBsAg显示两条多肽,分子量分别为23k和27k,与血源HBsAg的多肽成分相同,为HBsAg的两条特异性多肽。经PAGE银染色结果显示,纯化的HBsAg中杂蛋白含量符合疫苗制备要求。用上述方法提纯HBsAg,回收率可达44.1％以上。 将纯化的HBsAg吸附于氢氧化铝佐剂,免疫Balb/c小鼠,并与血源HBsAg对照,抗体半数阳转剂量(ED50)分别为0.501μg和0.832μg,说明基因工程HBsAg的免疫原性似优于血源HBsAg。
The purification of HBsAg in MT-5 cells culture supernatant was carried out by three different steps including ultrafiltration, precipitation by polyethylene glycol ( PEG ) and three-step ultracentrifugations. The purified HBsAg was composed of two bands in silver stained SDS-PAGE, their molecular weights being 23k and 27k respectively which were identical to those of human plasma HBsAg. There was no other protein band in the gel. The purity of purified HBsAg accorded with the requirement of hepatitis B vaccine. Up to 44.1% of starting HBsAg was recovered at the end of the purification process.Recombinant HBsAg was absorbed to Al(OH)3. The results of immu-nogenicity in Balb/c mice showed that ED50 of cellular HBsAg was 0.501 μg, but that of human plasma HBsAg was 0.832μg. The immunogenicity of cellular HBsAg was some what better than that of human plasma HBsAg.
出处
《病毒学报》
CAS
CSCD
北大核心
1990年第3期205-209,共5页
Chinese Journal of Virology
