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左润秋等人源性 Fab 段噬菌体抗体库的构建及抗人β1 AR 抗体克隆的筛选第 4 期· 192 ·
人源性 Fab 段噬菌体抗体库的构建及抗人β1 AR 抗体克隆的筛选
左润秋赵树进王捷张宏斌邱健向定成(广州军区总医院,广州 510010)
中国图书分类号 R392 11 文献标识码 A 文章编号 1000 484X(2005) 04 0291 05
[摘要] 目的:构建人源性 Fab 段噬菌体抗体库并筛选抗人β1 AR 抗体克隆。方法:通过 RT PCR 从抗人β1 AR 抗体阳
性的扩张性心肌病(DCM) 患者的外周血淋巴细胞中扩增出人 IgG重链 Fd 段和κ、λ两轻链基因片段,将其克隆入噬粒 pComb3
中,构建人源性 Fab 段噬菌体抗体库,并利用噬菌体表面显示技术,以人β1 肾上腺素受体细胞外第二环 26 肽(β1 AREC Ⅱ) 为抗
原肽,对此抗体库进行淘筛富集。结果:成功地构建了库容量为 1 4 × 106 的人源性 Fab 段噬菌体抗体库,从此抗体库中筛选到
了特异性抗人β1 AR Fab 段噬菌体抗体阳性克隆。结论:利用噬菌体抗体库技术可以获得表达抗人β1 AR 抗体的重组克隆。
[关键词] β1 肾上腺素能受体;扩张性心肌病;噬菌体抗体库;噬菌体表面显示技术;Fab 段抗体
Construction of Fab fragment phage antibody library and selection of clones ex
pressing anti humanβ1 AR antibody
ZUO Run Qiu , ZHAO Shu Jin , WANG Jie , ZHANG Hong Bin , QIU Jian , XIANG Ding Cheng. Guangzhou Military
General Hospital , Guangzhou 510010 , China
[ Abstract] Objective :To construct a human Fab phage antibody library and to obtain some binant clones which can express Fab
fragment antibody againstβ1 adrenergic receptor. Methods :Fd heavy chain gene andκ、λlight chains gene of IgG obtained by RT PCR from
peripheral lymphocytes of DCMpatients whose antiβ1 AR antibodies are present were cloned into pComb3 vector and the human