2001 Fiscal Year Final Research Report Summary
Study of the pancreas specific glycopeptides for diagnosis of the pancreas cancer
| Project/Area Number |
11671201
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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 |
Digestive surgery
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| Research Institution | Hirosaki University |
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Principal Investigator |
YOSHIHARA Syuichi Hirosaki University, School of Medicine, Assistant Professor, 医学部, 講師 (00261454)
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| Co-Investigator(Kenkyū-buntansha) |
TOYOKI Yoshikazu Hirosaki University, School of Medicine, Instructor, 医学部・附属病院, 助手 (70301025)
SASAKI Mutsuo Hirosaki University, School of Medicine, Professor, 医学部, 教授 (10005077)
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| Project Period (FY) |
1999 – 2000
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| Keywords | human pancreatic juice / panceas cancer / tumor maker / glycopeptides |
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| Research Abstract |
Introduction
Prognosis of the pancreatic cancer is extremely poor. Poor prognosis of this disease partially consists in difficulty of early diagnosis for it. More sensitive tumor marker is necessary for early diagnosis of it.
Recently we have isolated fucose-rich glycopeptide from human pancreatic juice. AH glycopeptides are found to be free of sialic acid and galactosamine but contained unusually high content of fucose. Structures in these glycopeptides are very rare in the human body. In this study, we intend to make the relationship in these glycopeptides and pancreatic cancer clear.
Purification and immunization of these glycopeptides
Three glycopeptides (GP-2, GP-3 and GP4) were purified employing ion exchange chromatography and gel filtration. GP-2 had a tetraantennary structure with five L-fucose residues. We tried to make polyclonal antibody for this glycopeptides (GP-2). Antigen (Peptide-carrier conjugation) was dissolved in phosphate-buffered saline and emulsified in complete Freund's adjuvant. For antigen, 2 rabbits were immunized intradermally with 1 mg of antigen, with booster immunizations given intramuscularly every 2 weeks in an identical manner except that incomplete Freund's adjuvant was used. Anti-glycopeptide antibody responses in the serum were evaluated using an ELISA procedure.
Immunochemical Staining
Frozen sections (4-6 μm) of the pancreas were blocked with normal goat serum , and subsequently incubated with primary antibodies diluted. Secondary reagents were biotinylated goat-anti-rabbit Ig. Sections were counterstained with hematoxyline. Organelle of the acinar cell were partially stained.
Concentration of the serum
Serum concentrations were measured in normal control group and pancreatic cancer group by ELISA. Differentiations of serum concentration (173.4 vs. 146.4 μg/dl) in control group and cancer group were revealed.
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