Institut des Nanotechnologies de Lyon-INL, UMR CNRS 5270, Université de Lyon, 69134 Ecully, France.
Anal Biochem. 2010 May 1;400(1):10-8. doi: 10.1016/j.ab.2010.01.013. Epub 2010 Jan 15.
Protein microarray technology provides a useful approach for the simultaneous serodetection of various antibodies in low sample volumes. To implement functional protein microarrays, appropriate surface chemistry must be designed so that both the protein structure and the biological activity can be retained. In the current study, two surface chemistries for protein microarrays and immunofluorescent assays were developed. Glass slides were functionalized with N-hydroxysuccinimide (NHS) ester via a monofunctional silane or maleic anhydride-alt-methyl vinyl ether (MAMVE) copolymer to allow covalent grafting of histone proteins. Analytical performance of these microarrays was then evaluated for the detection of anti-histone autoantibodies present in the sera of patients suffering from a systemic autoimmune disease, namely systemic lupus erythematosus (SLE), and the results were compared with those of the classical enzyme-linked immunosorbent assay (ELISA) and Western blot. The detection limit of our MAMVE copolymer microarrays was 50-fold lower than that of the classical ELISA. Furthermore, 100-fold less volume of biological samples was required with these miniaturized immunoassays.
蛋白质微阵列技术为在小体积样本中同时检测各种抗体提供了一种有用的方法。为了实现功能性蛋白质微阵列，必须设计适当的表面化学，以保留蛋白质结构和生物活性。在本研究中，开发了两种蛋白质微阵列和免疫荧光分析的表面化学方法。通过单官能硅烷或马来酸酐-alt-甲基乙烯基醚（MAMVE）共聚物将 NHS 酯官能化到玻片上，以允许组蛋白蛋白的共价接枝。然后评估这些微阵列用于检测系统性自身免疫性疾病（如系统性红斑狼疮[SLE]）患者血清中存在的抗组蛋白自身抗体的分析性能，并将结果与经典酶联免疫吸附测定（ELISA）和 Western blot 进行比较。我们的 MAMVE 共聚物微阵列的检测限比经典 ELISA 低 50 倍。此外，这些微型免疫分析需要的生物样品体积减少了 100 倍。
