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针对禽流感H7N9血凝素蛋白的单克隆抗体的产生与特性鉴定

Generation and characterisation of monoclonal antibodies specific to avian influenza H7N9 haemagglutinin protein.

作者信息

Malik A, Mallajosyula Vva, Mishra N N, Arukha A P, Varadarajan R, Gupta S K

机构信息

Reproductive Cell Biology Laboratory, National Institute of Immunology, New Delhi, India.

Molecular Biophysics Unit, n Institute of Science, Bengaluru, Karnataka, India.

出版信息

Indian J Med Microbiol. 2016 Oct-Dec;34(4):489-494. doi: 10.4103/0255-0857.195366.

Abstract

INTRODUCTION

Emerging virulent strains of influenza virus pose a serious public health threat with potential pandemic consequences. A novel avian influenza virus, H7N9, breached the species barrier from infected domestic poultry to humans in 2013 in China. Since then, it has caused numerous infections in humans with a close contact to poultry.

MATERIALS AND METHODS

In this study, we describe the preliminary characterisation of five murine monoclonal antibodies (MAbs) developed against recombinant haemagglutinin (rHA) protein of avian H7N9 A/Anhui/1/2013 virus by their Western blot and enzyme-linked immunosorbent assay (ELISA) reactivity and binding affinity.

RESULTS

Of the five MAbs, four were highly specific to H7N9 HA and did not show any cross-reactivity in ELISA with rHA protein from pandemic as well as seasonal H1N1, H2N2, H3N2, H5N1 and influenza virus B (B/Brisbane/60/2008). However, one of the MAbs, MA-24, in addition to HA protein of H7N9 also reacted strongly with HA protein of H3N2 and weakly with HA of pandemic and seasonal H1N1 and H2N2. All the five MAbs also reacted with H7N9 rHA in Western blot. The MAbs bound H7N9 rHA with an equilibrium dissociation constant (KD) ranging between 0.14 and 25.20 nM, indicating their high affinity to HA.

CONCLUSIONS

These antibodies may be useful in developing diagnostic tools for the detection of influenza H7N9 virus infections.

摘要

引言

新出现的流感病毒毒株对公众健康构成严重威胁,有可能引发大流行。2013年,一种新型禽流感病毒H7N9在中国突破了从感染家禽到人类的物种屏障。从那时起,它已在与家禽密切接触的人群中引发了大量感染。

材料与方法

在本研究中,我们描述了针对禽H7N9 A/Anhui/1/2013病毒重组血凝素(rHA)蛋白开发的五种鼠单克隆抗体(MAb)的初步特性,通过蛋白质印迹法和酶联免疫吸附测定(ELISA)检测它们的反应性和结合亲和力。

结果

在这五种单克隆抗体中,四种对H7N9 HA具有高度特异性,在ELISA中与大流行以及季节性H1N1、H2N2、H3N2、H5N1和B型流感病毒(B/Brisbane/60/2008)的rHA蛋白均未显示任何交叉反应。然而,其中一种单克隆抗体MA-24,除了与H7N9的HA蛋白反应外,还与H3N2的HA蛋白强烈反应,与大流行和季节性H1N1以及H2N2的HA蛋白弱反应。所有五种单克隆抗体在蛋白质印迹法中也与H7N9 rHA反应。这些单克隆抗体与H7N9 rHA结合的平衡解离常数(KD)在0.14至25.20 nM之间,表明它们对HA具有高亲和力。

结论

这些抗体可能有助于开发用于检测H7N9流感病毒感染的诊断工具。

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