1998 Fiscal Year Final Research Report Summary
Development of Immunoassay Method for Bio-active protein on the Basis of New Precise Labeling Techniques.
| Project/Area Number |
09470528
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Laboratory medicine
|
|---|
| Research Institution | KYUSHU UNIVERSITY |
|---|
Principal Investigator |
ZAITSU Kiyoshi KYUSHU UNIVERSITY,FAC.PHARM.SCI., PROFESSOR, 薬学部, 教授 (70091329)
|
|---|
| 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)
|
|---|
| Project Period (FY) |
1997 – 1998
|
| Keywords | immunoassay / labeling / peroxidase / microperoxidase / heme / insulin / transferrin / protecting group |
|---|
| 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.
|
