Liu Jianping, Eddings Mark A, Miles Adam R, Bukasov Rostislav, Gale Bruce K, Shumaker-Parry Jennifer S
Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, USA.
Anal Chem. 2009 Jun 1;81(11):4296-301. doi: 10.1021/ac900181f.
Surface Plasmon Resonance Microscopy (SPRM) is a promising label-free analytical tool for the real-time study of biomolecule interactions in a microarray format. However, flow cell design and microarray fabrication have hindered throughput and limited applications of SPRM. Here we report the integration of a microfluidic flow cell array (MFCA) with SPRM enabling in situ microarray fabrication and multichannel analysis of biomolecule probe-target interactions. We demonstrate the use of the MFCA for delivery of sample solutions with continuous flow in 24 channels in parallel for rapid microarray creation and binding analysis while using SPRM for real-time monitoring of these processes. Label-free measurement of antibody-antibody interactions demonstrates the capabilities of the integrated MFCA-SPRM system and establishes the first steps of the development of a high-throughput, label-free immunogenicity assay. After in situ probe antibody immobilization, target antibody binding was monitored in real time in 24 channels simultaneously. The limit of detection for this particular antibody pair is 80 ng/mL which is approximately 6 times lower than the industry recommended immunogenicity assay detection limit. The integrated MFCA-SPRM system is a powerful and versatile combination for a range of array-based analyses, including biomarker screening and drug discovery.
表面等离子体共振显微镜（SPRM）是一种很有前景的无标记分析工具，可用于以微阵列形式实时研究生物分子相互作用。然而，流动池设计和微阵列制造阻碍了SPRM的通量并限制了其应用。在此，我们报告了一种微流控流动池阵列（MFCA）与SPRM的集成，实现了原位微阵列制造以及生物分子探针 - 靶标相互作用的多通道分析。我们展示了使用MFCA在24个通道中并行连续流动输送样品溶液，用于快速创建微阵列和结合分析，同时使用SPRM实时监测这些过程。无标记测量抗体 - 抗体相互作用证明了集成的MFCA - SPRM系统的能力，并为高通量、无标记免疫原性分析的开发迈出了第一步。在原位固定探针抗体后，同时在24个通道中实时监测靶标抗体的结合。这种特定抗体对的检测限为80 ng/mL，比行业推荐的免疫原性分析检测限低约6倍。集成的MFCA - SPRM系统是用于一系列基于阵列分析（包括生物标志物筛选和药物发现）的强大且通用的组合。
