重组人源性抗HBsAg Fab抗体的特性分析  

Characterization analysis of recombinant humanized anti-HBsAg Fab antibody

作　　者：邓宁[1] 向军俭[1] 陈文吟[2] 王珣章[3] 粟宽源[2] 

机构地区：[1]暨南大学生物工程学系,广东广州510632 [2]解放军第458医院传染病研究中心,广东广州510602 [3]中山大学生物防治国家重点实验室,生物医药中心

出　　处：《暨南大学学报（自然科学与医学版）》2004年第5期656-660,共5页Journal of Jinan University(Natural Science & Medicine Edition)

摘　　要：　通过噬菌体展示技术从含高滴度抗乙肝表面抗原(HBsAg)抗体的人外周血淋巴细胞获得抗HBsAgFab抗体的轻、重链基因.将Fab的轻、重链基因分别整合到巴斯德毕赤酵母(Pichiapastoris)GS115菌株的染色体上,成功构建了高效分泌表达抗HBsAgFab抗体的酵母工程菌.对酵母表达的重组Fab抗体进行了纯化,并对其相对分子质量、糖基化以及抗原结合能力等特性进行了分析,结果显示重组酵母分泌表达的重组人源性抗HBsAgFab是一个相对分子质量为50000左右的低糖基化糖蛋白.1mg重组Fab抗体相当于40U的抗乙肝表面抗原抗体(20U/mg).表明重组Fab抗体具有较强的结合HBsAg的能力.The genes of anti-HBsAg Fab antibody were obtained from human peripheral blood lymphocytes of volunteer with high antibody titer to HBsAg by using of phage display technology. Here a high-level expression system to directly produce anti-HBsAg Fab antibody was established in Pichia pastoris. Which was achieved by co-integration of the genes encoding the heavy and light chains both under the genome of the yeast cells. The recombinant Fab antibody has been purified, and the molecular, glycosylation and affinity activity have been analized. The results demonstrated that the recombinant Fab antibody is a glycoprotein and it could sufficiently neutralize the HBsAg.

关 键 词：FAB抗体 亲和纯化 毕赤酵母 

分 类 号：R392.11[医药卫生—免疫学] Q786[医药卫生—基础医学]