High-Resolution Analysis of Antibodies to Post-Translational Modifications Using Peptide Nanosensor Microarrays.
High-Resolution Analysis of Antibodies to Post-Translational Modifications Using Peptide Nanosensor Microarrays.
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DOI：
10.1021/acsnano.6b03786
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发表时间：
2016-12-27
期刊：
影响因子：
17.1
通讯作者：
Wang SX
中科院分区：
文献类型：
--
作者：
Lee JR;Haddon DJ;Gupta N;Price JV;Credo GM;Diep VK;Kim K;Hall DA;Baechler EC;Petri M;Varma M;Utz PJ;Wang SX
自身抗体是红斑狼疮等自身免疫性疾病的标志，有可能被用作多种疾病的生物标志物，包括免疫缺陷、传染病和癌症。需要更精确地检测针对特定目标的抗体，以提高对此类疾病的诊断。在这里，我们报告了可重复使用的肽微阵列的发展，基于巨磁阻（GMR）纳米传感器，优化为灵敏地检测磁性纳米颗粒标记，用于检测具有单个翻译后修饰氨基酸的抗体。我们还开发了一种化学再生方案，可进行高重复性的多重分析，从而大大降低了实验成本。此外，我们发现直接在纳米传感器上合成的肽的灵敏度大约是直接点化肽的两倍。可重复使用的肽纳米传感器微阵列能够以高分辨率和灵敏度精确检测自身抗体，并有望研究抗体介导的对自身抗原、疫苗和病原体来源抗原的免疫反应，以及其他基本的肽-蛋白相互作用。
Autoantibodies are a hallmark of autoimmune diseases such as lupus and have the potential to be used as biomarkers for diverse diseases, including immunodeficiency, infectious disease, and cancer. More precise detection of antibodies to specific targets is needed to improve diagnosis of such diseases. Here, we report the development of reusable peptide microarrays, based on giant magnetoresistive (GMR) nanosensors optimized for sensitively detecting magnetic nanoparticle labels, for the detection of antibodies with a resolution of a single post-translationally modified amino acid. We have also developed a chemical regeneration scheme to perform multiplex assays with a high level of reproducibility, resulting in greatly reduced experimental costs. In addition, we show that peptides synthesized directly on the nanosensors are approximately two times more sensitive than directly spotted peptides. Reusable peptide nanosensor microarrays enable precise detection of autoantibodies with high resolution and sensitivity and show promise for investigating antibody-mediated immune responses to autoantigens, vaccines, and pathogen-derived antigens as well as other fundamental peptide–protein interactions.