Bio-MEMS를 기반으로 microfilter와 백금 전극이 내재되어 있는 microbiochip을 제작하였다. 우리는 이 chip으로 microbead에 indirect ELISA 방법으로 항원-항체를 반응시키고 전기 신호 검출 방법을 이용하여 반응 여부를 판단하였다. 이 때 신호 증폭을 위해 silver enhancer를 사용하였다. Chip 상에서 항원-항체 반응 조건을 최적화하기 위해 pH, temperature, flow time, flow rate, silver enhancer time을 결정하였다. 이렇게 최적화된 조건을 바탕으로 짧은 시간 안에 소량의 시료로 immunoassay를 성공적으로 수행할 수 있었다. 전기 신호 검출 방식을 사용함으로써 biosensor 장비의 소형화와 다중 시료 측정과 자동화를 biosensor에서 적용할 수 있는 가능성을 제시하였다.
This study presents the characterization of an integrated portable microfluidic electrical detection system for fast and low volume immunoassay using polystyrene microbead, which are used as immobilization surfaces. In our chip, a filtration method using the microbead was adopted for sample immobilization and immunogold silver staining(IGSS) was used to increase the electrical signal. The chip is composed of an inexpensive and biocompatible Polydimethylsiloxane(PDMS) layer and Pyrex glass substrate. Platinum microelectrodes for electric signal detection were fabricated on the substrate and microchannel and pillar-type microfilters were formed in the PDMS layer. With a fabricated chip, we reacted antigen and antibody according to the procedures. Then, silver enhancer was injected to increase the size of nanogold particles tagged with the second antibody. As a result, microbeads were connected to each other and formed an electrical bridge between microelectrodes. Resistance measured through the electrodes showed a difference of two orders of magnitude between specific and nonspecific immuno-reactions. The detection limit was 10 ng/ml. The developed immunoassay chip reduced the total analysis time from 3 hours to 50 min. Fast and low-volume biochemical analysis has been successfully achieved with the developed microfilter and immuno-sensor chip, which is integrated to the microfluidic system.
