| Project/Area Number |
09470528
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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 |
Laboratory medicine
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
ZAITSU Kiyoshi KYUSHU UNIVERSITY,FAC.PHARM.SCI., PROFESSOR, 薬学部, 教授 (70091329)
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| Co-Investigator(Kenkyū-buntansha) |
HAMASE Kenji KYUSHU UNIVERSITY,FAC.PHARM.SCI., Instructor (Jyosyu), 薬学部, 助手 (10284522)
OHBA Yoshihito KYUSHU UNIVERSITY,FAC.PHARM.SCI., Instructor (Jyosyu), 薬学部, 助手 (50274478)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 1998: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1997: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | immunoassay / labeling / peroxidase / microperoxidase / heme / insulin / transferrin / protecting group / パルオキシダーゼ / 免疫測定法 / 標識法 / タンパク質 |
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| Research Abstract |
A basic concept in immunoassays for restricted introduction of Label molecules onto protein is originated to establish firm basis of universal and superior labeling methods.
1) Mono-horseradish peroxidase(HRP)-labeled insulin afforded higher sensitivity in detecting insulin than the diHRP-insulin(our previous data). A single or plural numbers of HRP was labeled on transferrin (Tr), a medium size protein. The sensitivity obtained with mono-HRP-Tf was higher than that with multi-HRP- labeled Tf. This indicates that multi-HRP-labeling cause decrease of antigen- antibody binding. In practice, we developed a sensitive determination method for urinary Tf by using mono-HRP-Tf without special pre-treatment of urine sample.
2) Gly(Al)- and Lys(B29)-microperoxidase-11(HUP)-insulin showed almost the same ability in enzyme-immunoassay of insulin. When hemin, small label, was used for chemiluminescence label, the sensitivity attained with random-hemin-labeled- insulin was higher than that with mono-hemin-insulin. As little as 20 fmol insulin could be determined.
3) Gly(Al)- Phe(Bl)- and Lys(B29)-HUP labeled insulin were successfully prepared by using 3,4,5,6-tetrahydrophthalic anhydride as a reversible amino- protecting reagent. Thus, we can obtain mono-label-insulins anytime.
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