2003 Fiscal Year Final Research Report Summary
Development of On-Site Environmental Monitoring Microsystems Using Miniaturized Ultrathin Slab Gel Eletrophoresis
| Project/Area Number |
13124201
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
|
| Allocation Type | Single-year Grants |
|---|
| Review Section |
Science and Engineering
|
|---|
| Research Institution | Gunma University |
|---|
Principal Investigator |
UMEMURA Tomonari Gunma University, Department of Chemistry, Research Assistant, 工学部, 助手 (10312901)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
UCHIYAMA Kenji Tokyo Metropolitan University, Department of Chemistry, Research Assistant, 大学院・理学研究科, 助手 (00321901)
KORANAGA Takashi Tokyo Metropolitan University, Department of Chemistry, Professor, 大学院・理学研究科, 教授 (30124788)
TSUNODA Kin-ichi Gunma University, Department of Chemistry, Professor, 工学部, 教授 (30175468)
ODAKE Tamao Gunma University, Department of Chemistry, Lecturer, 工学部, 講師 (10301128)
|
|---|
| Project Period (FY) |
2001 – 2003
|
| Keywords | Two-dimensional electrophoresis / Protein / Isoelectric focusing / Optical waveguide / Interface / Air monitoring / Diesel exhaust particular / Micro TOF-Mass spectrometer |
|---|
| Research Abstract |
1-1. Miniaturized slab gel electrophoresis was developed and experimental conditions for the miniaturized gel were optimized. Miniaturized slab gel of 30 mm wide, 30 mm long, and 0.1 mm thick was applied to two-dimensional electrophoresis (2-DE), and feasibility of high performance separation of proteins were investigated.
1-2. Multifilament string-gel (MFS-Gel) was proposed as an isoelectric focusing gel (first dimension gel of 2-DE). The MFS-Gel could provide 1 mm thin, blow concentration gel down to 2.5%, which resulted in applicability to proteins of higher molecular weight that 200 kDa.
1-3. Fast 2-De using the MFS-Gel was developed. Colored proteins were used to evaluate performance of the isoelectric focusing system, and focusing field could be applied up to 430 V/cm, and the migration time could be reduced from 2 hours to 30 min. Practical samples such as E. Coli and chicken skeletal muscle extract were separated in 2 hours in the first dirnension and in 1 hour in the second dimension, which was at least three times faster than the conventional 2-DE.
1-4. Optical waveguide detection system using the evanescent wave was developed for microchip analysis and interfacial substances were successfully detected.
2-1. Microchip-device has been developed and optimized in order to measure for atmospheric NO_2 with high sensitivity. The detection limit was approximately 1 ppb in wind tunnel.
2-2. Pre-treatment microchip was developed for eluting PAHs from diesel exhaust particle (DEP). PAHs in DEP were trapped into silica gel beads, which were packed into the microchip, and extracted into chloroform. The recovery of this pre-treatment microchip was same as bulk scale methods.
2-3. MS interface has been developed. This interface has monolith structure in the capillary, and investigated the capacity to resist pressure between vacuum bath and liquid delivery system. As a result of connecting this interface and micro liquid delivery system, flow rate has reduced below 1 μL/min.
|
