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脂蛋白捕获 ELISA 法用于灵敏检测两亲性生物标志物。

Lipoprotein capture ELISA method for the sensitive detection of amphiphilic biomarkers.

机构信息

Physical Chemistry and Applied Spectroscopy, Chemistry Division, Los Alamos National Laboratory, Los Alamos, NM, United States.

Physical Chemistry and Applied Spectroscopy, Chemistry Division, Los Alamos National Laboratory, Los Alamos, NM, United States.

出版信息

Anal Biochem. 2022 Sep 1;652:114747. doi: 10.1016/j.ab.2022.114747. Epub 2022 May 27.

Abstract

Enzyme-linked immunosorbent assays (ELISAs) are widely employed for the detection of protein targets due to their ease of use, sensitivity, and potential for high-throughput analyses. However, the use of ELISAs to detect non-protein targets such as lipids and amphiphiles is complicated by the physical properties of these molecules, which affects their association with functional surfaces and recognition ligands. Here, we developed a unique lipoprotein capture ELISA in which the natural association between lipoproteins and amphiphilic molecules facilitates detection of the target biomarker in a physiologically relevant conformation. An assay to detect the glycolipid lipoarabinomannan (LAM), a cell membrane component and virulence factor associated with Mycobacterial infections, was developed as a proof of concept.

摘要

酶联免疫吸附测定(ELISA)因其使用简便、灵敏度高以及高通量分析的潜力而被广泛用于蛋白质靶标的检测。然而,由于这些分子的物理性质,ELISA 用于检测非蛋白质靶标(如脂质和两亲分子)变得复杂,这会影响它们与功能表面和识别配体的结合。在这里,我们开发了一种独特的脂蛋白捕获 ELISA,其中脂蛋白与两亲分子之间的天然关联促进了以生理相关构象检测目标生物标志物。作为概念验证,开发了一种检测糖脂脂阿拉伯甘露聚糖(LAM)的测定方法,LAM 是与分枝杆菌感染相关的细胞膜成分和毒力因子。

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