Sensitive immuno-detection using enzyme immobilized in nanoporous silica = 나노다공성 실리카를 이용한 고 민감도의 면역 진단

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we developed a GOx immobilized nanoporous MCF silica and magnetic bead based sandwich immuno-detection system. Taking advantage of the unique structural characteristics of the nanoporous MCF silica, the large pore volume and surface area, and mesopores with large diameter and uniform sizes, we successfully immobilized the large enzyme, GOx into the pores using cross-linking agent, also modified the surface of the MCF silica with antibodies. As a result, the MCF-GOx-Ab was used to detecte antigens captured by magnetic bead-Ab through sandwich approach. At optimized conditions, 107beads/mL of magnetic beads, and 10min of catalytic reaction time, using GOx immobilized in MCF silica it could detect model antigens, human IgG from 100pM to 100nM with less nonspecific binding to chicken IgG., and it showed about 20times higher sensitivity than conventional immunoassay using Ab-GOx conjugate antibodies made in our lab. To improve the sensitivity of this immuno detection, several factors still need to be optimized, for instance, the concentration of magnetic beads, the enzyme loading percentage, the quantity of antibodies linked to the surface of MCF silica and the concentrations of the substrates for catalytic reaction. Also different enzymes like HRP (horseradish peroxidase), or ALP (alkaline phosphatase) could be tried to develop a sensitive immunoprobe. As a new immunosensing technology, it must have profound potential not only in detection of various antigens but pathogens as well.
Advisors
Kim, Hak-Sungresearcher김학성researcher
Description
한국과학기술원 : 생명과학과,
Publisher
한국과학기술원
Issue Date
2006
Identifier
260019/325007  / 020044323
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생명과학과, 2006.8, [ iv, 25 p. ]

Keywords

감지 신호 높임; 항원-항체 결합; 민감도; 자기성 구슬; 나노다공성 MCF 실리카; 면역진단; sensitivity; antigen-antibody interaction; signal amplification; immuno-detection; nanoporous MCF silica; Magnetic bead

URI
http://hdl.handle.net/10203/28078
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=260019&flag=dissertation
Appears in Collection
BS-Theses_Master(석사논문)
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