ReScan, a Multiplex Diagnostic Pipeline, Pans Human Sera for SARS-CoV-2 Antigens.

ReScan, a Multiplex Diagnostic Pipeline, Pans Human Sera for SARS-CoV-2 Antigens.
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DOI:
10.1016/j.xcrm.2020.100123
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发表时间:
2020-10-20
期刊:
Cell reports. Medicine
影响因子:
--
通讯作者:
Wilson MR
Wilson MR
中科院分区:
其他
文献类型:
--
作者:
Zamecnik CR;Rajan JV;Yamauchi KA;Mann SA;Loudermilk RP;Sowa GM;Zorn KC;Alvarenga BD;Gaebler C;Caskey M;Stone M;Norris PJ;Gu W;Chiu CY;Ng D;Byrnes JR;Zhou XX;Wells JA;Robbiani DF;Nussenzweig MC;DeRisi JL;Wilson MR

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对SARS-CoV-2感染的血清学反应的全面了解对于病理生理学洞察和诊断发展都很重要。在这里,我们生成了一种泛人类冠状病毒可编程噬菌体展示分析,以进行由98份COVID-19患者血清富集的冠状病毒抗原的蛋白质组全谱分析。接下来,我们使用ReScan,这是一种有效隔离表达最具免疫原性肽的噬菌体的方法,并使用声学液体处理将它们打印到纸基微阵列上,该方法分离并鉴定了9种候选抗原,其中8种来自用于SARS-CoV-2血清学检测的两种蛋白质:刺突蛋白和核衣壳蛋白。在适用于临床实验室环境的高通量测定中部署后,这些抗原显示出比整个蛋白质组更高的特异性。这项概念验证研究表明,ReScan将广泛适用于其他缺乏有效生物标志物的新兴传染病或自身免疫性疾病,从而实现从抗原发现到使用一种重组蛋白源进行诊断的无缝管道。ReScan是一个完整的蛋白质组筛选,用于分离和鉴定血清学检测靶标COVID-19血清中线性肽的抗体结合刺突蛋白和核衣壳蛋白快速工作流程,将生物标志物无缝转化为功能诊断多重线性S和N SARS-CoV-2肽可提高诊断特异性在抗原未知的情况下,快速血清学测定法的开发仍然具有挑战性,并且在诊断过程中特别重要。流行病Zamecnik等人使用ReScan,它结合了纸基微阵列和可编程噬菌体展示,筛选和分离SARS-CoV-2抗体诊断的最具免疫原性的肽。
Comprehensive understanding of the serological response to SARS-CoV-2 infection is important for both pathophysiologic insight and diagnostic development. Here, we generate a pan-human coronavirus programmable phage display assay to perform proteome-wide profiling of coronavirus antigens enriched by 98 COVID-19 patient sera. Next, we use ReScan, a method to efficiently sequester phage expressing the most immunogenic peptides and print them onto paper-based microarrays using acoustic liquid handling, which isolates and identifies nine candidate antigens, eight of which are derived from the two proteins used for SARS-CoV-2 serologic assays: spike and nucleocapsid proteins. After deployment in a high-throughput assay amenable to clinical lab settings, these antigens show improved specificity over a whole protein panel. This proof-of-concept study demonstrates that ReScan will have broad applicability for other emerging infectious diseases or autoimmune diseases that lack a valid biomarker, enabling a seamless pipeline from antigen discovery to diagnostic using one recombinant protein source. ReScan is a whole proteome screen to isolate and identify serologic assay targets Antibodies to linear peptides in COVID-19 sera bind spike and nucleocapsid proteins Rapid workflow that seamlessly translates biomarkers into a functional diagnostic Multiplexing linear S and N SARS-CoV-2 peptides can increase diagnostic specificity Rapid serologic assay development remains challenging in scenarios in which antigens are unknown and is particularly crucial during a pandemic. Zamecnik et al. use ReScan, which combines paper-based microarrays and programmable phage display, to screen for and isolate the most immunogenic peptides for SARS-CoV-2 antibody diagnostics.
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