Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus
Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus
复制标题
DOI：
10.1016/j.jviromet.2004.04.009
复制
发表时间：
2004-09-01
影响因子：
3.1
通讯作者：
Plummer, F
中科院分区：
文献类型：
--
作者：
Berry, JD;Jones, S;Plummer, F
目前全球都需要阐明SARS冠状病毒（SARS-CoV）表达的保护性抗原。因此，迫切需要能够识别SARS冠状病毒特异性抗原的单克隆抗体试剂。在这份报告中，开发和免疫化学特性的一组鼠单克隆抗体（单克隆抗体）对SARS冠状病毒的基础上，其特异性，结合要求，和生物活性。用高度纯化的病毒作为包被抗原，通过ELISA进行初步筛选，结果选择了103个针对SARS病毒的mAb。随后的筛选步骤将该组减少至17种IgG mAb。如在Western免疫印迹中所见，单个mAb F26 G15对核蛋白具有特异性，而其他五种mAb与刺突蛋白反应。这些刺突特异性mAb中的两个证明了体外中和SARS-CoV的能力，而另外四个Western免疫印迹阴性mAb也中和了病毒。这些单克隆抗体的诊断开发的效用被证明。来自恢复期SARS患者的抗体，而不是正常人血清，也显示出特异性竞争mAb与整个SARS-CoV的结合。这些研究强调了使用标准化检测和试剂的重要性。这些单克隆抗体将有助于SARS冠状病毒诊断试验的发展，致病机制的研究和疫苗的开发。(C)2004 Elsevier B. V.保留所有权利。
There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development. (C) 2004 Elsevier B.V. All rights reserved.