Drobin Kimi, Nilsson Peter, Schwenk Jochen M
Science for Life Laboratory Stockholm, School of Biotechnology, KTH-Royal Institute of Technology, Stockholm, Sweden.
Methods Mol Biol. 2013;1023:137-45. doi: 10.1007/978-1-4614-7209-4_8.
Alongside the increasing availability of affinity reagents, antibody microarrays have become a powerful tool to screen for target proteins in complex samples. Applying directly labeled samples onto arrays instead of using sandwich assays offers an approach to facilitate a systematic, high-throughput, and flexible exploration of protein profiles in body fluids such as serum or plasma. As an alternative to planar arrays, a system based on color-coded beads for the creation of antibody arrays in suspension has become available to offer a microtiter plate-based option for screening larger number of samples with variable sets of capture reagents. A procedure was established for analyzing biotinylated samples without the necessity to remove excess labeling substance. We have shown that this assay system allows detecting proteins down into lower pico-molar and higher pg/ml levels with dynamic ranges over three orders of magnitude. Presently, this workflow enables the profiling of 384 samples for up to 384 proteins per assay.
随着亲和试剂的可得性不断提高，抗体微阵列已成为筛选复杂样品中靶蛋白的强大工具。将直接标记的样品应用于阵列而非使用夹心测定法，为在血清或血浆等体液中系统、高通量且灵活地探索蛋白质谱提供了一种方法。作为平面阵列的替代方案，一种基于颜色编码珠子的系统可用于在悬浮液中创建抗体阵列，为使用可变捕获试剂集筛选大量样品提供了基于微孔板的选择。建立了一种分析生物素化样品的方法，无需去除过量的标记物质。我们已经表明，该检测系统能够检测低至皮摩尔级和高至皮克/毫升水平的蛋白质，动态范围超过三个数量级。目前，该工作流程每次测定可对384个样品中的多达384种蛋白质进行分析。
