融合多肽表位用于 SARS-CoV-2 抗体检测的高级多重血清学测试

IF 3.8 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY ACS Bio & Med Chem Au Pub Date : 2023-08-30 DOI:10.1021/acsbiomedchemau.3c00010
Ali H. Aldoukhi, Panayiotis Bilalis, Dana M. Alhattab, Alexander U. Valle-Pérez, Hepi H. Susapto, Rosario Pérez-Pedroza, Emiliano Backhoff-García, Sarah M. Alsawaf, Salwa Alshehri, Hattan Boshah, Abdulelah A. Alrashoudi, Waleed A. Aljabr, Manal Alaamery, May Alrashed, Rana M. Hasanato, Raed A. Farzan, Roua A. Alsubki, Manola Moretti, Malak S. Abedalthagafi and Charlotte A. E. Hauser*, 
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引用次数: 0

摘要

悲惨的 COVID-19 大流行病在全世界已累计出现 6.55 亿例病例,死亡人数超过 660 万,似乎终于要结束了。即便如此,严重急性呼吸系统综合症冠状病毒 2(SARS-CoV-2)的新变种仍在不断出现,其严重程度无法事先预测。这关系到公共卫生的维护和稳定,因为可能会出现免疫逃避和传播能力增强的情况。因此,继续监测普通人群对 SARS-CoV-2 的抗体反应至关重要。作为聚合酶链反应测试的补充,多重免疫测定是一种优雅的工具,它同时使用单个蛋白质或肽抗原,具有很高的灵敏度和特异性。为了进一步提高 SARS-CoV-2 抗体检测的灵敏度和准确性,我们开发了一种先进的基于多肽的血清学多重检测方法,使用的抗原融合肽表位来自尖峰蛋白和核壳蛋白。多肽表位与已报道的 SARS-CoV-2 抗体之间的分子对接模拟验证了抗体检测所选表位的重要性。与全长蛋白质相比,肽的修饰和合成更容易、更快,因此可以以更具成本效益的方式使用。我们合成了三种不同的融合表位肽(FEPs),并用酶联免疫吸附试验(ELISA)进行了测试。共使用了 145 份血清样本,其中 110 份来自 COVID-19 患者的 COVID-19 血清样本,35 份来自疫情爆发前未感染 COVID-19 的阴性对照血清样本。有趣的是,我们的数据表明,FEP 抗原的灵敏度、特异性和准确性均高于单肽表位或单肽表位混合物。我们的 FEP 概念不仅可用于检测 SARS-CoV-2 还可用于检测其他各种病原体,适用于不同的多重免疫测定。经过大幅改进的多肽血清学检测方法可支持商业化护理点检测方法(如横向流动检测方法)的开发。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection

The tragic COVID-19 pandemic, which has seen a total of 655 million cases worldwide and a death toll of over 6.6 million seems finally tailing off. Even so, new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to arise, the severity of which cannot be predicted in advance. This is concerning for the maintenance and stability of public health, since immune evasion and increased transmissibility may arise. Therefore, it is crucial to continue monitoring antibody responses to SARS-CoV-2 in the general population. As a complement to polymerase chain reaction tests, multiplex immunoassays are elegant tools that use individual protein or peptide antigens simultaneously to provide a high level of sensitivity and specificity. To further improve these aspects of SARS-CoV-2 antibody detection, as well as accuracy, we have developed an advanced serological peptide-based multiplex assay using antigen-fused peptide epitopes derived from both the spike and the nucleocapsid proteins. The significance of the epitopes selected for antibody detection has been verified by in silico molecular docking simulations between the peptide epitopes and reported SARS-CoV-2 antibodies. Peptides can be more easily and quickly modified and synthesized than full length proteins and can, therefore, be used in a more cost-effective manner. Three different fusion-epitope peptides (FEPs) were synthesized and tested by enzyme-linked immunosorbent assay (ELISA). A total of 145 blood serum samples were used, compromising 110 COVID-19 serum samples from COVID-19 patients and 35 negative control serum samples taken from COVID-19-free individuals before the outbreak. Interestingly, our data demonstrate that the sensitivity, specificity, and accuracy of the results for the FEP antigens are higher than for single peptide epitopes or mixtures of single peptide epitopes. Our FEP concept can be applied to different multiplex immunoassays testing not only for SARS-CoV-2 but also for various other pathogens. A significantly improved peptide-based serological assay may support the development of commercial point-of-care tests, such as lateral-flow-assays.

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ACS Bio & Med Chem Au
ACS Bio & Med Chem Au 药物、生物、化学-
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期刊介绍: ACS Bio & Med Chem Au is a broad scope open access journal which publishes short letters comprehensive articles reviews and perspectives in all aspects of biological and medicinal chemistry. Studies providing fundamental insights or describing novel syntheses as well as clinical or other applications-based work are welcomed.This broad scope includes experimental and theoretical studies on the chemical physical mechanistic and/or structural basis of biological or cell function in all domains of life. It encompasses the fields of chemical biology synthetic biology disease biology cell biology agriculture and food natural products research nucleic acid biology neuroscience structural biology and biophysics.The journal publishes studies that pertain to a broad range of medicinal chemistry including compound design and optimization biological evaluation molecular mechanistic understanding of drug delivery and drug delivery systems imaging agents and pharmacology and translational science of both small and large bioactive molecules. Novel computational cheminformatics and structural studies for the identification (or structure-activity relationship analysis) of bioactive molecules ligands and their targets are also welcome. The journal will consider computational studies applying established computational methods but only in combination with novel and original experimental data (e.g. in cases where new compounds have been designed and tested).Also included in the scope of the journal are articles relating to infectious diseases research on pathogens host-pathogen interactions therapeutics diagnostics vaccines drug-delivery systems and other biomedical technology development pertaining to infectious diseases.
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