Chip-based immunoassays.
Chip-based immunoassays.
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DOI：
10.1007/978-1-62703-029-8_21
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发表时间：
2013
影响因子：
--
通讯作者：
A. Apori;A. Herr
中科院分区：
文献类型：
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作者：
A. Apori;A. Herr
微流控免疫分析技术在速度、自动化和便携性方面优于台式金标准对应物。具体地，芯片上免疫减法是用于报告蛋白质天然迁移率和结合特异性的快速均相免疫测定。通过基于尺寸的排阻过滤器从电泳检测中去除抗体结合的靶蛋白，而未结合的非靶蛋白能够通过过滤器进行下游检测。通过制造离散图案化的聚丙烯酰胺（PA）凝胶区域在芯片上实现免疫减影。此外，PA凝胶区域用于定义芯片上样品制备区域，用于蛋白富集、荧光标记和免疫减影前的抗体-靶标结合。在这里，我们描述了免疫减影装置的制造技术，以及用于确定目标蛋白的流动性和结合特异性的复杂的生物样品，包括脑脊液内的电泳分析协议。
Microfluidic immunoassay techniques offer advantages in speed, automation, and portability over bench-top gold standard counterparts. In particular, on-chip immunosubtraction is a rapid homogeneous immunoassay used for reporting both protein native mobility and binding specificity. Immunosubtraction is performed by removing antibody-bound target proteins from electrophoretic detection via a size-based exclusion filter, while unbound nontarget proteins are able to pass through the filter for downstream detection. Immunosubtraction is achieved on-chip by fabrication of discrete patterned polyacrylamide (PA) gel regions. Additionally, PA gel regions are used to define on-chip sample preparation regions for protein enrichment, fluorescent labeling, and antibody-target binding prior to immunosubtraction. Here we describe the immunosubtraction device fabrication technique as well as the electrophoretic assay protocol for determining target protein mobility and binding specificity within complex biological samples including cerebrospinal fluid.