2006 Fiscal Year Final Research Report Summary
Changes on glycosylation and serum levels of acute phase protein in severe viral infection.
| Project/Area Number |
17580280
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Clinical veterinary science
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| Research Institution | Yamaguchi University |
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Principal Investigator |
IWATA Hiroyuki Yamaguchi University, Faculty of Agriculture, Professor, 農学部, 教授 (40193750)
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| Co-Investigator(Kenkyū-buntansha) |
MAEDA Ken Yamaguchi University, Faculty of Agriculture, Associate professor, 農学部, 助教授 (90284273)
OKUDA Masaru Yamaguchi University, Faculty of Agriculture, Associate professor, 農学部, 助教授 (10325243)
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| Project Period (FY) |
2005 – 2006
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| Keywords | acute phase protein / glycan chain / virus / infectious disease / cancer |
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| Research Abstract |
The purpose of this study is to clarify a significance of α_1-acid glycoprotein (AGP) as a model of glycobiology of acute phase proteins in inflammation, cancer and viral infection.
1. Preparation of AGP in animals: Feline AGP (fAGP) was purified from ascites by ammonium sulfate precipitation, acid precipitation, ion-exchanged column chromatography, and gel filtration. The purified protein showed a broad band with an M. W. of 40-50 kDa and high solubilities against ammonium sulfate and acid solution with a lower pI than 3.7, indicating that the protein was confirmed as feline AGP.
2. Preparation of monoclonal antibodies(Mabs) against feline AGP: Mabs were prepared as the ordinary method. In result, we obtained 8 Mabs of which isotypes (L chains) were IgG1(λ),IgG2b(λ) and IgA(κ). On Western blot analyses, these Mabs recognized fAGP in non-reducing conditions, but not in reducing conditions. In addition, competitive ELISA showed these Mabs recognized the same or overlaped epitope.
3. Lectin affinity: Feline AGP had an affinity against ConA, DSA, SAA and UEA-1, having di-antenary and tri-antenary structures, sialic acid and fucose in glycan chains.
4. Preparation of antiserum and determination of fAGP: Antiserum was prepared in a rabbit.
Ouchterlony test showd a continuous immunoprecipitin line against fAGP and feline serum. Direct binding ELISA using the antiserum with determination range of 0.02 to 0.25 pig/m1 showed serum fAGP levels of 1.37± 1.147 mg/ml.
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