Autoantigen microarrays for multiplex characterization of autoantibody responses
Autoantigen microarrays for multiplex characterization of autoantibody responses
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DOI：
10.1038/nm0302-295
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发表时间：
2002-03-01
期刊：
影响因子：
82.9
通讯作者：
Utz, PJ
中科院分区：
文献类型：
--
作者：
Robinson, WH;DiGennaro, C;Utz, PJ
我们构建了小型化的自身抗原阵列，使用亚微升数量的临床样本对针对结构多样的自身抗原的自身抗体反应进行大规模的多重表征。自身抗原微阵列是通过使用机器人阵列仪将数百种蛋白质、肽和其他生物分子附着到衍生化载玻片表面来生产的。将阵列与患者血清一起孵育，并使用光谱可分辨的荧光标记来检测与阵列上的特异性自身抗原结合的自身抗体。我们描述并表征了包含8种不同人类自身免疫性疾病（包括系统性红斑狼疮和类风湿性关节炎）中主要自身抗原的阵列。这代表了将这种技术应用于多种人类疾病血清的第一份报告，并且将使得能够验证识别自身抗原的抗体的检测，所述自身抗原包括蛋白质、肽、酶复合物、核糖核蛋白复合物、DNA和免疫后修饰的抗原。自身抗原微阵列是研究自身抗体反应的特异性和发病机制以及鉴定和定义人类自身免疫性疾病中相关自身抗原的有力工具。
We constructed miniaturized autoantigen arrays to perform large-scale multiplex characterization of autoantibody responses directed against structurally diverse autoantigens, using submicroliter quantities of clinical samples. Autoantigen microarrays were produced by attaching hundreds of proteins, peptides and other biomolecules to the surface of derivatized glass slides using a robotic arrayer. Arrays were incubated with patient serum, and spectrally resolvable fluorescent labels were used to detect autoantibody binding to specific autoantigens on the array. We describe and characterize arrays containing the major autoantigens in eight distinct human autoimmune diseases, including systemic lupus erythematosus and rheumatoid arthritis. This represents the first report of application of such technology to multiple human disease sera, and will enable validated detection of antibodies recognizing autoantigens including proteins, peptides, enzyme complexes, ribonucleoprotein complexes, DNA and post-translationally modified antigens. Autoantigen microarrays represent a powerful tool to study the specificity and pathogenesis of autoantibody responses, and to identify and define relevant autoantigens in human autoimmune diseases.