| Budget Amount *help |
¥41,990,000 (Direct Cost: ¥32,300,000、Indirect Cost: ¥9,690,000)
Fiscal Year 2018: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
Fiscal Year 2017: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
Fiscal Year 2016: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
Fiscal Year 2015: ¥17,030,000 (Direct Cost: ¥13,100,000、Indirect Cost: ¥3,930,000)
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| Outline of Final Research Achievements |
The purpose of this research was to develop a high-speed method for selection of synthetic binding proteins (SBPs) and quantitative detection of the corresponding target proteins at single-molecule resolution. First, we modified our previous high-speed method of screening functional polypeptides (transcription-translation coupled with association of puromycin linker [TRAP] display; JACS, 2013) for SBPs selection. Then, using the improved TRAP display, we demonstrated successful selection of high-affinity (nM level) SBPs against model target proteins, EGFR1 and HER2. Finally, we applied the modified method to obtain several SBPs against low-copy-number proteins and to detect a target protein immobilized on a glass plate using a fluorescently labeled SBP. The improved version of the TRAP display for SBP selection and single-molecule detection of target proteins with the obtained SBPs could become a basic high-precision technology for protein quantification at single-molecule resolution.
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