摘要
目的 :纯化酵母发酵产生的重组人源性抗HBsAgFab抗体 ,建立稳定、适于生产应用的纯化工艺。方法 :以原核表达的重组人源性抗HBsscFv免疫BALB/c小鼠 ,经杂交瘤技术制备大量分泌mAb的杂交瘤细胞株 14F7。将该细胞株注入小鼠腹腔制备mAb腹水 ,经辛酸沉淀纯化后 ,制备亲和层析柱 ,纯化酵母发酵产生的重组人源性抗HBsAgFab抗体。结果 :用抗scFvmAb亲和层析柱纯化的重组人源性抗HBsAgFab的纯度可达 95 %以上 ,回收率可达 70 %～ 85 %。结论 :人源性抗HBsscFv制备mAb作为亲和层析的介质 ,可有效地从酵母发酵上清中纯化重组人源性抗HBsAgFab,适用于大规模生产重组人源性抗HBsAgFab。
AIM: To establish a steady purification method for producing recombinant humanized anti-HBsAg Fab from yeast fermentation supernatant. METHODS: Humanized anti-HBs scFv was used to immunize BALB/c mice to obtain anti-scFv monoclonal antibody (mAb) which was used to purify recombinant humanized anti-HBsAg Fab from yeast fermantation supernatant by affinity chromatography. RESULTS: The purity of purified recombinant anti-HBsAg Fab was above 95% and its recovery rate was about 75%-85%. CONCLUSION: An efficient affinity chromatography suitable for purification of recombinant humanized anti-HBsAg Fab in large-scale was established.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第5期575-577,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
广州市科技局重点项目资助 (No .2 0 0 3J1 C0 1 71 )
国家自然科学基金项目资助 (No .40 3760 30 )
