Development of a High-Throughput Mass Spectrometry-Based SARS-CoV-2 Immunoassay.
Development of a High-Throughput Mass Spectrometry-Based SARS-CoV-2 Immunoassay.
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DOI：
10.1021/acs.analchem.3c02421
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发表时间：
2023-12
影响因子：
7.4
通讯作者：
Jie Sun;Jong Hee Song;Mary K Danielson;Nathan D Colley;Alia Thomas;David Hambly;J. C. Barnes;Michael L. Gross
中科院分区：
文献类型：
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作者：
Jie Sun;Jong Hee Song;Mary K Danielson;Nathan D Colley;Alia Thomas;David Hambly;J. C. Barnes;Michael L. Gross
Covid-19 大流行的严重影响凸显了对快速、可靠和高通量感染诊断方法的需求。目前的分析方法，无论是现场检测还是集中检测，都无法满足患者友好型检测、高要求和结果可靠性的要求。在这里，我们提出了一种两点分离按需诊断策略，该策略使用激光解吸/电离飞行时间质谱（LDI-TOF MS）并采用稳定但可裂解的离子探针作为质量报告基因。使用该报告基因可以实现超灵敏、可中断、可存储、可恢复和高通量的按需检测。我们描述了这一概念的演示，其中我们（i）设计和合成激光可裂解报告基因（DTPA），（ii）将报告基因缀合到抗体上并验证缀合物的功能，（iii）以良好的周转和高灵敏度检测缀合的报告基因，（iv）使用激光可裂解内标进行定量分析，以及（v）识别含有刺突蛋白的阴性和阳性样品。该方案具有出色的灵敏度（amol 适用于 SARS-CoV-2 Spike S1 亚基抗体），无需任何扩增。该策略也适用于除 SARS-CoV-2 之外的其他疾病抗原的检测。
The serious impact of the Covid-19 pandemic underscores the need for rapid, reliable, and high-throughput diagnosis methods for infection. Current analytical methods, either point-of-care or centralized detection, are not able to satisfy the requirements of patient-friendly testing, high demand, and reliability of results. Here, we propose a two-point separation on-demand diagnostic strategy that uses laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS) and adopts a stable yet cleavable ionic probe as a mass reporter. The use of this reporter enables ultrasensitive, interruptible, storable, restorable, and high-throughput on-demand detection. We describe a demonstration of the concept whereby we (i) design and synthesize a laser-cleavable reporter (DTPA), (ii) conjugate the reporter onto an antibody and verify the function of the conjugate, (iii) detect with good turnaround and high sensitivity the conjugated reporter, (iv) analyze quantitatively by using a laser-cleavable internal standard, and (v) identify negative and positive samples containing the spike protein. The protocol has excellent sensitivity (amol for the SARS-CoV-2 Spike S1 subunit antibody) without any amplification. This strategy is also applicable for the detection of other disease antigens besides SARS-CoV-2.