通过简便的侧向流动免疫测定法快速、特异性地检测氧化型低密度脂蛋白/β2糖蛋白I复合物。

Rapid and specific detection of oxidized LDL/β2GPI complexes via facile lateral flow immunoassay.

作者信息

Tan Xian Wen, Takenaka Fumiaki, Takekawa Hironori, Mastuura Eiji

机构信息

Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.

Collaborative Research Center (OMIC), Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.

出版信息

Heliyon. 2020 Jun 8;6(6):e04114. doi: 10.1016/j.heliyon.2020.e04114. eCollection 2020 Jun.

DOI:10.1016/j.heliyon.2020.e04114

PMID:32551380

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7287255/

Abstract

β2-Glycoprotein I (β2GPI) forms indissociable complex with oxidized LDL (oxLDL) into proatherogenic oxLDL/β2GPI complex through a specific ligand known as 7-ketocholesteryl-9-carboxynonanoate (oxLig-1). Recent discoveries have demonstrated the atherogenicity of these complexes in patients of both systemic and non-systemic autoimmune diseases. Hence, serological level of oxLDL/β2GPI complexes may represent one crucial clinical parameter for disease prognosis of atherosclerosis-related diseases. Herein, we established a simple, specific and rapid gold nanoparticle (GNP) based lateral flow immunoassay (LFIA) to quantify oxLDL/β2GPI complexes from test samples. Specificities of hybridoma cell-derived monoclonal antibodies against antigen, optimal conditions for conjugation of antibody with GNP, and sensitivity of oxLDL/β2GPI LFIA in comparison to an ELISA-based detection method were assessed accordingly. The established oxLDL/β2GPI LFIA was capable of detecting oxLDL/β2GPI specifically without interference from autoantibodies and solitary components of oxLDL/β2GPI present in test samples. A significant correlation (R > 0.8) was also obtained with the oxLDL/β2GPI LFIA when compared to the ELISA-based detection. On the whole, the oxLDL/β2GPI LFIA remains advantageous over the oxLDL/β2GPI ELISA. The unnecessary washing step, short developmental and analytical time support facile and rapid detection of oxLDL/β2GPI as opposed to the laborious ELISA system.

摘要

β2糖蛋白I（β2GPI）通过一种名为7-酮胆固醇基-9-羧基壬酸酯（oxLig-1）的特定配体与氧化低密度脂蛋白（oxLDL）形成不可解离的复合物，进而形成促动脉粥样硬化的oxLDL/β2GPI复合物。最近的研究发现表明，这些复合物在系统性和非系统性自身免疫性疾病患者中具有动脉粥样硬化性。因此，oxLDL/β2GPI复合物的血清学水平可能是动脉粥样硬化相关疾病疾病预后的一个关键临床参数。在此，我们建立了一种基于金纳米颗粒（GNP）的简单、特异且快速的侧向流动免疫分析（LFIA）方法，用于定量检测测试样品中的oxLDL/β2GPI复合物。相应地评估了杂交瘤细胞衍生的抗抗原单克隆抗体的特异性、抗体与GNP偶联的最佳条件，以及与基于酶联免疫吸附测定（ELISA）的检测方法相比，oxLDL/β2GPI LFIA的灵敏度。所建立的oxLDL/β2GPI LFIA能够特异性检测oxLDL/β2GPI，不受测试样品中自身抗体和oxLDL/β2GPI单独成分的干扰。与基于ELISA的检测方法相比，oxLDL/β2GPI LFIA也获得了显著的相关性（R>0.8）。总体而言，oxLDL/β2GPI LFIA相对于oxLDL/β2GPI ELISA仍具有优势。与繁琐费力的ELISA系统相比，无需洗涤步骤、检测和分析时间短，支持对oxLDL/β2GPI进行简便快速的检测。