Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense M, 5230, Denmark.
Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, Aarhus C, 8000, Denmark.
Sci Rep. 2017 May 15;7(1):1925. doi: 10.1038/s41598-017-02214-0.
To date, there are multiple assays developed that detect and quantify antibodies in biofluids. Nevertheless, there is still a lack of simple approaches that specifically detect autoimmune antibodies to double-stranded DNA. Herein we investigate the potential of novel nucleic acid complexes as targets for these antibodies. This is done in a simple, rapid and specific immunofluorescence assay. Specifically, employing 3D nanostructures (DNA origami), we present a new approach in the detection and study of human antibodies to DNA. We demonstrate the detection of anti-DNA antibodies that are characteristic of systemic lupus erythematosus, a chronic autoimmune disease with multiple manifestations. We tested the most potent non-covalent pairs of DNA and fluorescent dyes. Several complexes showed specific recognition of autoimmune antibodies in human samples of lupus patients using a simple one-step immunofluorescence method. This makes the novel assay developed herein a promising tool for research and point-of-care monitoring of anti-DNA antibodies. Using this method, we for the first time experimentally confirm that the disease-specific autoimmune antibodies are sensitive to the 3D structure of nucleic acids and not only to the nucleotide sequence, as was previously thought.
迄今为止,已经开发出多种检测生物体液中抗体的方法,并能对其进行定量分析。然而,目前仍然缺乏简单的方法来特异性检测针对双链 DNA 的自身抗体。在此,我们研究了新型核酸复合物作为这些抗体的靶标的潜力。这是通过一种简单、快速和特异性的免疫荧光检测法来实现的。具体来说,我们采用 3D 纳米结构(DNA 折纸术),提出了一种用于检测和研究人类针对 DNA 的抗体的新方法。我们证明了可以检测出系统性红斑狼疮(一种具有多种表现的慢性自身免疫性疾病)特征性的抗 DNA 抗体。我们测试了最有效的非共价 DNA 和荧光染料对。使用简单的一步免疫荧光法,几种复合物在狼疮患者的人类样本中显示出对自身抗体的特异性识别。这使得本文开发的新检测方法成为研究和即时检测抗 DNA 抗体的有前途的工具。通过该方法,我们首次从实验上证实,疾病特异性自身抗体对核酸的 3D 结构敏感,而不仅仅是对核苷酸序列敏感,这与之前的观点不同。