Microbead based micro total analysis system for Hepatitis C detection

마이크로비드를 이용한 초소형 C형 간염 검출 시스템의 제작

  • Sim, Tae-Seok (School of Electrical Engineering and Computer Science, Seoul National University) ;
  • Lee, Bo-Rahm (School of Chemical and Biological Engineering, Seoul National University) ;
  • Lee, Sang-Myung (School of Chemical and Biological Engineering, Seoul National University) ;
  • Kim, Min-Soo (School of Electrical Engineering and Computer Science, Seoul National University) ;
  • Lee, Yoon-Sik (School of Chemical and Biological Engineering, Seoul National University) ;
  • Kim, Byung-Gee (School of Chemical and Biological Engineering, Seoul National University) ;
  • Kim, Yong-Kweon (School of Electrical Engineering and Computer Science, Seoul National University)
  • 심태석 (서울대학교 전기.컴퓨터공학부) ;
  • 이보람 (서울대학교 화학생물공학부) ;
  • 이상명 (서울대학교 화학생물공학부) ;
  • 김민수 (서울대학교 전기.컴퓨터공학부) ;
  • 이윤식 (서울대학교 화학생물공학부) ;
  • 김병기 (서울대학교 화학생물공학부) ;
  • 김용권 (서울대학교 전기.컴퓨터공학부)
  • Published : 2006.07.12

Abstract

This paper describes a micro total analysis system ($\mu$ TAS) for detecting and digesting the target protein which includes a bead based temperature controllable microchip and computer based controllers for temperature and valve actuation. We firstly combined the temperature control function with a bead based microchip and realized the on-chip sequential reactions using two kinds of beads. The PEG-grafted bead, on which RNA aptamer was immobilized, was used for capturing and releasing the target protein. The target protein can be chosen by the type of RNA aptamer. In this paper, we used the RNA aptamer of HCV replicase. The trypsin coated bead was used for digesting the released protein prior to the matrix assisted laser desorption ionization time of flight mass spectrometer (MALDI TOF MS). Heat is applied for release of the captured protein binding on the bead, thermal denaturation and trypsin digestion. PDMS microchannel and PDMS micro pneumatic valves were also combined for the small volume liquid handling. The entire procedures for the detection and the digestion of the target protein were successfully carried out on a microchip without any other chemical treatment or off-chip handling using $20\;{\mu}l$ protein mixture within 20 min. We could acquire six matched peaks (7% sequence coverage) of HCV replicase.

Keywords