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利用 Faradaic Shotgun Tagging 对复杂介质中的目标蛋白生物标志物进行定量分析。

A Quantification of Target Protein Biomarkers in Complex Media by Faradaic Shotgun Tagging.

机构信息

Department of Chemistry, University of Oxford South Parks Road, Oxford OX1 3QZ, U.K.

Department of Clinical Biochemistry, John Radcliffe Hospital Oxford University Hospitals NHS Foundation Trust, Oxford OX3 9DU, U.K.

出版信息

Anal Chem. 2022 Feb 8;94(5):2375-2382. doi: 10.1021/acs.analchem.1c03695. Epub 2022 Jan 27.

Abstract

The progressive emergence of protein biomarkers promises a revolution in the healthcare industry and a shift of focus from disease management to much earlier intervention. Here, we introduce a facile shotgun tagging of ensemble proteins in clinically relevant media prior to specific target capture at antibody-modified electrodes. This facilitates a convenient voltammetric quantification of markers down to sub-pg/mL levels and across several orders of concentration. A translation of the methodology to an automated microfluidic platform enables marker quantification from 25 μL of sample in less than 15 min, demonstrated here with a simultaneous assaying of CRP and cardiac troponin I (cTnI). The assays show a good correlation with a standard immunoassay when applied to real patient serum samples. The platform is simple, generic, highly sensitive and requires no secondary labeling/binding or amplification.

摘要

蛋白质生物标志物的逐渐出现有望给医疗保健行业带来一场革命,并将重点从疾病管理转移到更早的干预。在这里,我们介绍了一种在抗体修饰电极进行特定目标捕获之前,在临床相关介质中对蛋白质进行简便的鸟枪法标记的方法。这使得可以方便地通过伏安法对标记物进行定量分析,检测下限可达亚 pg/mL 水平,并跨越几个浓度数量级。该方法的转化为自动化微流控平台提供了便利,可在不到 15 分钟的时间内从 25μL 的样品中定量标记物,这里同时检测了 C-反应蛋白(CRP)和心肌肌钙蛋白 I(cTnI)。该测定方法应用于真实的患者血清样本时,与标准免疫测定法具有良好的相关性。该平台简单、通用、高度灵敏,并且不需要二次标记/结合或放大。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b9e/9082491/b9e2b90fd959/ac1c03695_0008.jpg

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