| Project/Area Number |
01480331
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Digestive surgery
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| Research Institution | Saitama Medical School |
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Principal Investigator |
ISHIDA Kiyoshi Saitama Medical School, Faculty of Medicine, Professor, 医学部, 教授 (80049767)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAJIMA Hiroshi Tokyo Medical and Dental University, Medical Research Institute, Department of B, 難治疾患研究所, 教授 (70013810)
YAMAZAKI Masahiko Saitama Medical School, Faculty of Medecine, Research Associate, 医学部, 助手 (60220323)
KURIHARA Shigekatsu Saitama Medical School, Faculty of Medicine, Lecturer, 医学部, 講師 (90192038)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1990: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1989: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Bilirubin-albumin / Monoclinal antibody / Enzyme-linked immunosorbent assay / Enzymeーlinked immunosorbent assay / ビリルビン / 単クロ-ン抗体 / ELISA / anti-oxidant |
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| Research Abstract |
A new anti-bilirubin monoclinal antibody has been prepared and characterized. Using modified MBS (metamaleimidobenzoyl N-hydroxysuccinimide ester) method, haptenic unconjugated bilirubin-IXalpha (UCB) covalently coupled to bovine serum albumin (BSA) retaining its intramolecular hydrogen bonds and three-dimensional structure. After somatic cell fusion and screeing assay, we obtained anti-bilirubin monoclonal antibodies which reacted with UCB but did not react with bilirubin diglucuronide or monoglucuronide.
To determine extremely low concentrations of bilirubin in human serum or urine, we utilized an antibody presently available (5M2) and another anti-bilirubin monoclinal antibody (24G7) which reacted with UCB as well as with conjugated bilirubin for enzyme-linked immunosorbent assay (ELISA). Bilirubin concentration in serm specimens obtained from many patients, determined using ELISA was compared with that determined by the conventional diazo method. When 5M2 was used in ELIZA, there was no discrepancy between bilirubin concentration determined by the two method in any sample. However, when 24G7 was used in ELISA, in the case of patients whose total urine volume was decreased, there was a significant discrepacy. This suggests the existence of a degraded bilirubin, substance "X", that is increased in the serum when any kind of stress occurs which will be excreted in the urine, In addition existence of substance "X" in the urine of healthy patients was observed.
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