Summary of Research Project Results under JSPS FY2002
"Research for the Future Program"

1.Research Institution Japan Advanced Institute of Science and Technology
2.Research Area Integrated Fields
3.Research Field Biomedical Observation and Control
4.Term of Project FY 1998 - FY 2002
5.Project Number 98I00401
6.Title of Project Creation of Generic Technology for Advanced Biosensors

7.Project Leader
Name Institution,Department Title of Position
Eiichi, Tamiya Japan Advanced Institute of Science and Technology, School of Materials Science Professor

8.Core Members

Name Institution,Department Title of Position
Kenji, Yokoyama National institute of Advanced Industrial Science and Technology, Laboratory of Advanced Bioelectronics Deputy Director
Kazuyoshi, Yano Tokyo University of Technology, Katayanagi Advanced Research Laboratories Associate Professor
Yasutaka, Morita Japan Advanced Institute of Science and Technology, School of Materials Science Assistant Professor

9.Summary of Research Results

  In this project, we developed the biosensor to be high stable as follows.
  We designed a sensor peptide based on structural change that takes place when MAPK (mytogen activated protein kinase) is phosphorylated by MAPKK (MAP kinase kinase) in the intracellular signal transduction. The imprinted polymer can strongly recognize an amine by the strong electrostatic interaction between the amino group of the amine and the sulfo group in the binding sites. This method provides an amine-recognizing polymer material by using a thiol derivative instead of the amine. We developed the construction of an artificial fructosyl amine dehydrogenase based on molecular imprinting technology and its application to the artificial enzyme sensor. We developed the molecularly imprinted catalyst (MIC). We have carried out to develop a high-though-put measurement system for the detection organic pollution (BOD) by using micro-chip immobilized luminous microorganisms. We made use of P19 as a model of stem cell and selected the peptides that bind with P19 from phage display library.
  We screened the DNA aptamer inhibiting Taq DNA polymerase with novel method combining systematic evolution of ligands by exponential enrichment(SELEX) and in silico evolution. By adopting an alkaline phosphatase-linked aptamer that binds to bile acids a new analytical method for the detection of bile acids has been developed. As a method to estimate the activities of G-protein-coupled receptor, we developed a method to detect phosphate ion (Pi) using europium (III) and time-resolved fluorescence spectrometry.
  A micro optical cantilever made of thin silicon dioxide film has been applied to DNA imaging and provided a 100 nm resolution fluorescence image. Imaging of single molecular fluorescence has been showed blinking and breaching phenomena that characterize the single molecular fluorescence.
  We have developed miniaturized biotechnology tools both of flow-type and array-format. As a flow-type device, a hydrophobic valve structure has newly fabricated with switchable surface modification that can change its hydrophilicity. Microchemical sensors and microactuators which would become key components in micro analysis systems were fabricated using micromachining techniques. These include an analysis system that imitates a mosquito, an on-line sensing system for ammonia and creatinine, an integrated microfluidic transport system using bubbles as active elements for pumps and valves. A portable surface plasmon resonance (SPR) sensor system consisting of a miniature SPR sensor chip Spreeta™, a hydrostatic pressure driven flow system using a Marionette bottle and a sample injector, was developed.

10.Key Words
(1)Biosensor   (2)Molecular recognition   (3)Combinatorial design
(4)Microchip technology   (5)Nanotechnology   (6)Biochip
(7)Environmental sensor   (8)Medical diagnostic sensor   (9)Cell-tissue technology