| Project/Area Number |
25630370
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
UEDA Hiroshi 東京工業大学, 資源化学研究所, 教授 (60232758)
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| Co-Investigator(Renkei-kenkyūsha) |
MATSUYAMA Yuki (OHMURO Yuki) 学術振興会RPD/神戸大学, 大学院工学研究科, 特命助教 (30571088)
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| Project Period (FY) |
2013-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2013: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | antibody / purification / affinity / avidity / immunoassay / 融合タンパク質 / 抗体 / 結合タンパク / 酵素 / リンカーペプチド / 抗体結合タンパク質 / アビディティー / リンカー工学 / 抗体工学 / 免疫測定 / 結合速度 / 解離速度 / 蛍光プローブ |
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| Outline of Final Research Achievements |
Two antibody binding domains from Protein A D (PA) and Protein G C1 (PG) were fused with a linker peptide with various lengths (DDAKK)n, n=0,2,4,6, to create a novel antibody binding protein PAxnPG. When their affinity to Fab and Fc of antibody was evaluated with a biosensor based on biolayer interferometry, the highest binding constant was obtained with n=4 for Fab, and n=6 for Fc, probably reflecting their length between two binding sites. Moreover, the obtained binding constants are several tens-fold higher than those with PA/PG alone. The utility of PAxPG as an antibody detection reagent was also suggested by immunoblot analysis using a fusion protein with chimeric human alkaline phosphatase (AP) with higher catalytic activity and stability than the parent APs from intestinal and placental origins.
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