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用于检测拉沙病毒核蛋白的单域抗体的筛选与鉴定

Selection and Characterization of Single-Domain Antibodies for Detection of Lassa Nucleoprotein.

作者信息

Anderson George P, Liu Jinny L, Shriver-Lake Lisa C, Goldman Ellen R

机构信息

Naval Research Laboratory, Center for Biomolecular Science and Engineering, Washington, DC 20375, USA.

出版信息

Antibodies (Basel). 2020 Dec 17;9(4):71. doi: 10.3390/antib9040071.

Abstract

Lassa virus is the etiologic agent of Lassa fever, an acute and often fatal illness endemic to West Africa. It is important to develop new reagents applicable either for the specific diagnosis or as improved therapeutics for the treatment of Lassa fever. Here, we describe the development and initial testing of llama-derived single-domain antibodies that are specific for the Lassa virus nucleoprotein. Four sequence families based on complementarity-determining region (CDR) homology were identified by phage-based enzyme-linked immunosorbent assays, however, the highest affinity clones all belonged to the same sequence family which possess a second disulfide bond between Framework 2 and CDR3. The affinity and thermal stability were evaluated for each clone. A MagPlex-based homogeneous sandwich immunoassay for Lassa virus-like particles was also demonstrated to show their potential for further development as diagnostic reagents.

摘要

拉沙病毒是拉沙热的病原体,拉沙热是一种在西非流行的急性且往往致命的疾病。开发适用于拉沙热特异性诊断或作为改进治疗方法的新试剂非常重要。在此,我们描述了针对拉沙病毒核蛋白的羊驼源单域抗体的开发和初步测试。通过基于噬菌体的酶联免疫吸附测定法鉴定了基于互补决定区(CDR)同源性的四个序列家族,然而,亲和力最高的克隆都属于同一个序列家族,该家族在框架2和CDR3之间具有第二个二硫键。对每个克隆的亲和力和热稳定性进行了评估。还展示了一种基于磁珠的针对拉沙病毒样颗粒的均相夹心免疫测定法,以表明它们作为诊断试剂进一步开发的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68de/7768477/7d233d24deaa/antibodies-09-00071-g0A1.jpg

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