Intel Corporation , Santa Clara, California 95052, United States.
Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California , Berkeley, California 94720, United States.
ACS Nano. 2016 Dec 27;10(12):10652-10660. doi: 10.1021/acsnano.6b03786. Epub 2016 Sep 20.
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.
自身抗体是狼疮等自身免疫性疾病的一个标志，有可能作为多种疾病的生物标志物，包括免疫缺陷、传染病和癌症。为了提高这些疾病的诊断准确性，需要更精确地检测针对特定靶标的抗体。在这里，我们报告了基于巨磁电阻（GMR）纳米传感器的可重复使用的肽微阵列的开发，该传感器经过优化，可灵敏地检测磁性纳米颗粒标记物，从而实现对单个翻译后修饰氨基酸的抗体的分辨率检测。我们还开发了一种化学再生方案，可进行具有高度重现性的多重分析，从而大大降低了实验成本。此外，我们表明，直接在纳米传感器上合成的肽比直接点样的肽敏感约两倍。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，并有望用于研究针对自身抗原、疫苗和病原体衍生抗原以及其他基本肽-蛋白相互作用的抗体介导的免疫反应。
