[从组合抗体文库中产生针对流感病毒的人重组抗体Fab片段]
[Generation of human recombinant antibody Fab fragment to influenza virus from combined antibody library].
作者信息
Yang G, Han F, Liang M
机构信息
State Key Lab for Molecular Virology and Genetic Engineering, Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, China.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2001 Jun;15(2):182-5.
OBJECTIVE
To acquire the recombinant human monoclonal antibodies to influenza virus which can be used in mucous membrane treatment to prevent the infection of virus in animal model in the future.
METHODS
Construct and pan human Fab antibody library to influenza virus Sydney strain (H3N2) from vaccinated donors by using phage display technology and secreted expression in E. Coli system. The positive Fab clones with neutralization activity to influenza virus were selected and characterized through HI test, IFA and VN test.
RESULTS
Separate two Fab clones, IV-2 and IV-6, appear positive responses in ELISA and IFA. These two clones can inhibit hemagglutination in HI test and can decrease viral titer by 30 and 20 times in viral neutralization assay, respectively.
CONCLUSION
We have got two clones of Fab fragments with neutralization activity to influenza virus Sydney strain (H3N2).
目的
获取重组人源抗流感病毒单克隆抗体，以便未来用于黏膜治疗，在动物模型中预防病毒感染。
方法
利用噬菌体展示技术，构建并淘选来自接种疫苗供体的针对流感病毒悉尼株（H3N2）的全人源Fab抗体文库，并在大肠杆菌系统中进行分泌表达。通过血凝抑制试验（HI试验）、间接免疫荧光法（IFA）和病毒中和试验（VN试验）筛选并鉴定对流感病毒具有中和活性的阳性Fab克隆。
结果
分离得到两个Fab克隆，IV-2和IV-6，在酶联免疫吸附测定（ELISA）和间接免疫荧光法（IFA）中呈现阳性反应。这两个克隆在血凝抑制试验中可抑制血凝，在病毒中和试验中分别可使病毒滴度降低30倍和20倍。
结论
我们获得了两个对流感病毒悉尼株（H3N2）具有中和活性的Fab片段克隆。
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