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通过神经节苷脂微阵列和表面等离子体共振成像检测血清中的多发性硬化症生物标志物

Detection of Multiple Sclerosis Biomarkers in Serum by Ganglioside Microarrays and Surface Plasmon Resonance Imaging.

作者信息

Malinick Alexander S, Lambert Alexander S, Stuart Daniel D, Li Bochao, Puente Ellie, Cheng Quan

机构信息

Department of Chemistry, University of California, Riverside, California 92521, United States.

Environmental Toxicology, University of California, Riverside, California 92521, United States.

出版信息

ACS Sens. 2020 Nov 25;5(11):3617-3626. doi: 10.1021/acssensors.0c01935. Epub 2020 Oct 28.

Abstract

Multiple sclerosis (MS) is an autoimmune disease that damages the myelin sheaths of nerve cells in the central nervous system. An individual suffering from MS produces increased levels of antibodies that target cell membrane components, such as phospholipids, gangliosides, and membrane proteins. Among them, anti-ganglioside antibodies are considered as important biomarkers to differentiate MS from other diseases that exhibit similar symptoms. We report here a label-free method for detecting a series of antibodies against gangliosides in serum by surface plasmon resonance imaging (SPRi) in combination with a carbohydrate microarray. The ganglioside array was fabricated with a plasmonically tuned, background-free biochip, and coated with a perfluorodecyltrichlorosilane (PFDTS) layer for antigen attachment as a self-assembled pseudo-myelin sheath. The chip was characterized with AFM and matrix-assisted laser desorption ionization mass spectrometry, demonstrating effective functionalization of the surface. SPRi measurements of patients' mimicking blood samples were conducted. A multiplexed detection of antibodies for anti-GT, anti-GM, and anti-GA in serum was demonstrated, with a working range of 1 to 100 ng/mL, suggesting that it is well suited for clinical assessment of antibody abnormality in MS patients. Statistical analyses, including PLS-DA and PCA show the array allows comprehensive characterization of cross reactivity patterns between the MS specific antibodies and can generate a wide range of information compared to traditional end point assays. This work uses PFDTS surface functionalization and enables direct MS biomarker detection in serum, offering a powerful alternative for MS assessment and potentially improved patient care.

摘要

多发性硬化症(MS)是一种自身免疫性疾病,会损害中枢神经系统中神经细胞的髓鞘。患有MS的个体产生的抗体水平会升高,这些抗体靶向细胞膜成分,如磷脂、神经节苷脂和膜蛋白。其中,抗神经节苷脂抗体被认为是区分MS与其他表现出相似症状疾病的重要生物标志物。我们在此报告一种无标记方法,通过表面等离子体共振成像(SPRi)结合碳水化合物微阵列来检测血清中一系列抗神经节苷脂抗体。神经节苷脂阵列是用经过等离子体调谐的无背景生物芯片制造的,并涂覆有全氟癸基三氯硅烷(PFDTS)层,用于作为自组装假髓鞘附着抗原。该芯片通过原子力显微镜(AFM)和基质辅助激光解吸电离质谱进行表征,证明了表面的有效功能化。对模拟患者血样进行了SPRi测量。结果表明能够对血清中的抗GT、抗GM和抗GA抗体进行多重检测,工作范围为1至100 ng/mL,这表明它非常适合对MS患者抗体异常进行临床评估。包括偏最小二乘判别分析(PLS-DA)和主成分分析(PCA)在内的统计分析表明,该阵列能够全面表征MS特异性抗体之间的交叉反应模式,与传统终点检测相比,可以生成更广泛的信息。这项工作采用PFDTS表面功能化,能够直接检测血清中的MS生物标志物,为MS评估提供了一种强大的替代方法,并有可能改善患者护理。

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