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2 6肽(β1 - ARECⅡ)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 4×10 6 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 -ARFab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。
Objective:To construct a human Fab phage antibody library and to obtain some recombinant clones which can express Fab fragment antibody against β 1-adrenergic receptor.Methods:Fd heavy chain gene and κ?λ light chains gene of IgG obtained by RT-PCR from peripheral lymphocytes of DCM patients whose anti β 1-AR antibodies are present were cloned into pComb3 vector and the human Fab phage antibody library was constructed.The library was panned by phage display technology with β 1-ARECⅡ as antigen.Results:A human Fab phage antibody library with 1.4×10 6 capability was constructed successfully,a positive clone against human β 1-AR was screened from the phage antibody library.Conclusion:The recombinant clones which express Fab antibody against β 1-AR can be obtained by phage display technology.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2005年第4期291-294,298,共5页
Chinese Journal of Immunology
