Fiber-optic microsphere-based antibody array for the analysis of inflammatory cytokines in saliva.
Fiber-optic microsphere-based antibody array for the analysis of inflammatory cytokines in saliva.
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DOI：
10.1021/ac802181j
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发表时间：
2009-03-15
影响因子：
7.4
通讯作者：
Walt, David R.
中科院分区：
文献类型：
--
作者：
Blicharz, Timothy M.;Siqueira, Walter L.;Helmerhorst, Eva J.;Oppenheim, Frank G.;Wexler, Philip J.;Little, Frederic F.;Walt, David R.
抗体微阵列已成为高通量蛋白质分析和候选生物标志物筛选的有用工具。我们在这里描述了一种多路微球抗体阵列的发展，能够同时测量十种炎症蛋白介质。细胞因子捕获微球是通过将目标细胞因子特异性单克隆抗体与荧光编码的3.1 μm聚合物微球共价偶联制备的。采用化学蚀刻方法，将含有约5万根直径3.1 μm的光纤束制成微孔，在微孔中沉积细胞因子捕获微球。微球随机分布在井中，以产生抗体阵列，用于进行多重夹心免疫测定。该阵列以浓度依赖性的方式对重组细胞因子溶液有特异性反应。该阵列还用于检测肺部炎症性疾病（如哮喘和慢性阻塞性肺疾病（COPD））患者唾液上清液中的内源性介质模式。这种阵列技术可能被证明是一种基于实验室的炎症疾病研究和诊断平台，它的小占地也可以集成到微流控盒中，用于即时检测。
Antibody microarrays have emerged as useful tools for high-throughput protein analysis and candidate biomarker screening. We describe here the development of a multiplexed microsphere-based antibody array capable of simultaneously measuring ten inflammatory protein mediators. Cytokine-capture microspheres were fabricated by covalently coupling monoclonal antibodies specific for cytokines of interest to fluorescently-encoded 3.1 μm polymer microspheres. An optical fiber bundle containing ∼50,000 individual 3.1 μm diameter fibers was chemically etched to create microwells in which cytokine-capture microspheres could be deposited. Microspheres were randomly distributed in the wells to produce an antibody array for performing a multiplexed sandwich immunoassay. The array responded specifically to recombinant cytokine solutions in a concentration-dependent fashion. The array was also used to examine endogenous mediator patterns in saliva supernatants from patients with pulmonary inflammatory diseases such as asthma and chronic obstructive pulmonary disease (COPD). This array technology may prove useful as a laboratory-based platform for inflammatory disease research and diagnostics, and its small footprint could also enable integration into a microfluidic cassette for use in point-of-care testing.
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