2004 Fiscal Year Final Research Report Summary
The investigation for high sensitive chemi luminescence method using nanoreactor as a marker
| Project/Area Number |
15350038
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KAMIDATE Tamio Hokkaido Univ., Grad.School of Eng., Prof., 大学院・工学研究科, 教授 (70185990)
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| Co-Investigator(Kenkyū-buntansha) |
TANI Hirofumi Hokkaido Univ., Grad.School of Eng., Asso.Prof., 大学院・工学研究科, 助教授 (10271644)
ISHIDA Akihiko Hokkaido Univ., Grad.School of Eng., Inst., 大学院・工学研究科, 助手 (20312382)
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| Project Period (FY) |
2003 – 2004
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| Keywords | liposome / nanoreactor / chemi luminescence / peroxidase / ruminol / immunoassay / homogentisic acid γ-lactone |
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| Research Abstract |
Liposomes encapsulated horseradish peroxidase(HRP) were prepared for applying the inner water phase in liposome to the nanoreactor for HRP-catalyzed chemiluminecence(CL) reaction. Phosphatidylcholine(PC), phosphatidylglycerol dimyristoyl(DMPG) and cholesterol were used as a component for liposome. The mole % of PC : DMPG : Chel was 8:1:1. The HRP-trapped liposome was prepared by extrusion method with polycarbonate filter. Homogentisic acid γ-lactone(HAL) and luminol were used as a CL reagent for the determination of horseradish peroxidase (HRP) encapsulated in liposomes. HRP was detected after the lysis of HRP-trapped liposomes with Triton X-100. CL response rate, detection limit and linear range of calibration curve for HRP in HAL CL were compared with those in ρ-iodophenol (ρ-IP) enhanced luminol CL. Maximal light emission in HAL CL appeared more rapidly compared to that in ρ-IP enhanced CL methods, thus resulting in remarkable reduction of CL measurement time. The detection limit for HRP in HAL CL was the same as that in ρ-IP enhanced lnminol CL. The linear range of calibration curve for HRP in HAL CL was improved by factors of 50 compared with that in ρ-IP enhanced luminol CL. From these results, it was found that HAL CL were superior to ρ-IP enhanced luminol CL for the determination of HRP encapsulated in liposomes.
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