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Nanocrystal-encoded fluorescent microbeads for proteomics: Antibody profiling and diagnostics of autoimmune diseases

机译：蛋白质组学的纳米晶体编码荧光微珠：自身免疫性疾病的抗体谱和诊断

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摘要

The first application of nanocrystal (NC)-encoded microbeads to clinical proteomics is demonstrated by multiplexed detection of circulating autoantibodies, markers of systemic sclerosis. Two-color complexes, consisting of NC-encoded, antigen-covered beads, anti-antigen antibody or clinical serum samples, and dye-tagged detecting antibodies, were observed using flow cytometry assays and on the surface of single beads. The results of flow cytometry assays correlated with the ELISA technique and provided clear discrimination between the sera samples of healthy donors and patients with autoimmune disease. Microbead fluorescence signals exhibited narrow distribution regardless of their surface antigen staining, without the need of any fluorescence compensation-a parameter determining the limit of sensitivity of flow cytometry assays. In single bead measurements, less than 30 dye-labeled antibodies interacting with the topoI-specific antibodies at the surface of a bead have been detected by the emission of dye excited through the FRET from NCs. In this format, the antibody-bead interaction reaction turns specifically the fluorescence signal from dye label off and on, additionally increasing autoantibody detection sensitivity.
机译：纳米晶体（NC）编码的微珠在临床蛋白质组学中的首次应用通过循环自身抗体（系统性硬化症的标志物）的多重检测得到证明。使用流式细胞仪检测并在单个珠子的表面上观察到了两种颜色的复合物，包括NC编码的，抗原覆盖的珠子，抗抗原抗体或临床血清样品以及染料标记的检测抗体。流式细胞仪检测的结果与ELISA技术相关，并提供了对健康供体和自身免疫性疾病患者血清样品的清晰区分。不论其表面抗原染色如何，微珠荧光信号均显示出狭窄的分布，而无需任何荧光补偿-该参数决定了流式细胞仪检测灵敏度的极限。在单珠测量中，通过从NCs中通过FRET激发的染料的发射，已经检测到不到30种与珠表面上的topoI特异性抗体相互作用的染料标记抗体。以这种格式，抗体-珠相互作用反应会特异性地打开和关闭来自染料标记的荧光信号，从而进一步提高了自身抗体检测的灵敏度。

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