Multiple enzyme linked immunosorbent assay system on a capillary-assembled microchip integrating valving and immuno-reaction functions.

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.