Henares Terence G, Funano Shun-ichi, Terabe Shigeru, Mizutani Fumio, Sekizawa Ryuichi, Hisamoto Hideaki
Graduate School of Material Science, University of Hyogo, 3-2-1 Kouto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
Anal Chim Acta. 2007 Apr 25;589(2):173-9. doi: 10.1016/j.aca.2007.02.060. Epub 2007 Mar 3.
Multiple enzyme linked immunosorbent assay (ELISA) chip is developed by using capillary-assembled microchip (CAs-CHIP) technique, which involves simple embedding of 2-3mm length of square capillaries possessing valving and immuno-reaction functions into the microchannels fabricated on a PDMS substrate. In contrast to the previously reported ELISA chips, our system enables not only the flexible design of the multi-ELISA chip required for many different diagnostic purposes, but also the valving operation required for a reliable analysis. Here, a thermo-responsive polymer-immobilized capillary was used together with a small Peltier device, as a valving part, and different antibody-immobilized capillaries were used as immuno-reaction part. Sample solution and detecting reagent solutions were sequentially introduced through the valving capillary, and the valve is closed to completely stop the solution flow inside the immuno-reaction capillaries and detected using thermal lens microscope (TLM). Different anti-IgGs (human, goat, chicken) were immobilized and used as ELISA parts of CAs-CHIP. Sequential introductions of the mixed IgG solution, mixed enzyme-antibody solution and substrate solution facilitated the multiple determinations of 0.1 ng mL(-1) IgGs (human, goat, chicken) with total analysis time of about 30 min. The valve-integrated multi-ELISA chip developed here can be applied for many different diagnostic purposes by using different immuno-reaction capillaries necessary for a specific clinical diagnostic application.
采用毛细管组装微芯片（CAs-CHIP）技术开发了多重酶联免疫吸附测定（ELISA）芯片，该技术涉及将长度为2-3mm、具有阀控和免疫反应功能的方形毛细管简单地嵌入到在聚二甲基硅氧烷（PDMS）基板上制造的微通道中。与先前报道的ELISA芯片相比，我们的系统不仅能够灵活设计用于多种不同诊断目的的多重ELISA芯片，还能实现可靠分析所需的阀控操作。在这里，使用热响应聚合物固定化毛细管与一个小型珀耳帖装置作为阀控部件，不同的抗体固定化毛细管作为免疫反应部件。样品溶液和检测试剂溶液通过阀控毛细管依次引入，然后关闭阀门以完全停止免疫反应毛细管内的溶液流动，并使用热透镜显微镜（TLM）进行检测。固定化不同的抗IgG（人、山羊、鸡）并用作CAs-CHIP的ELISA部件。依次引入混合IgG溶液、混合酶抗体溶液和底物溶液，可实现对0.1 ng mL(-1) IgG（人、山羊、鸡）的多重测定，总分析时间约为30分钟。这里开发的集成阀控多重ELISA芯片可通过使用特定临床诊断应用所需的不同免疫反应毛细管，应用于多种不同的诊断目的。
