Significance of Bilirubin as Anti-Oxidant in Gastrointestinal Surgery

胆红素作为抗氧化剂在胃肠手术中的意义

• 批准号: 01480331
• 负责人: ISHIDA Kiyoshi
• 金额: $ 4.29万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01480331/
• 关键词: 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

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.

制备并鉴定了一种新的抗胆红素单抗。利用修饰的甲基亚胺酰苯甲酰基n-羟基琥珀酰亚胺酯（MBS）方法，将半偶联胆红素- ix - α (UCB)与牛血清白蛋白（BSA）共价偶联，保留了其分子内氢键和三维结构。经过体细胞融合和筛选实验，我们获得了抗胆红素单克隆抗体，该抗体与UCB反应，而与双脲类胆红素或单脲类胆红素不反应。为了确定人血清或尿液中极低浓度的胆红素，我们使用了目前可用的抗体（5M2）和另一种抗胆红素单抗（24G7），该抗体与UCB和偶联胆红素反应，用于酶联免疫吸附试验（ELISA）。用ELISA法与传统重氮法测定的胆红素浓度进行了比较。在ELIZA中使用5M2时，两种方法测定的任何样品的胆红素浓度均无差异。然而，在ELISA中使用24G7时，在总尿量减少的患者中，存在明显的差异。这表明存在一种降解的胆红素，即物质“X”，当任何一种应激发生时，它在血清中增加，并通过尿液排出。此外，在健康患者的尿液中也观察到存在物质“X”。

项目成果

岡村 維摩,山崎 順彦,山口 登喜夫,菰田 泰夫,中島 煕: "Antiーbilirubin monoclonal antibody.III.Preparation and properties of monoclonal antibodies to unconjugated bilirubinーIXα" Biochimica et Biophysica Acta.

Ima Okamura、Yoshihiko Yamazaki、Tokio Yamaguchi、Yasuo Komoda、Hiroshi Nakajima：“抗胆红素单克隆抗体。III.非结合胆红素-IXα单克隆抗体的制备和特性”Biochimica et Biophysica Acta。

岡村 維摩、山崎 順彦、山口 登喜夫、菰田 泰夫、中島 熈: "Antiーbilirubin monoclonal antibody. III.Preparation and properties of monoclonal antibodies to unconjugated bilirubinーIXα" Biochimica et Biophysica Acta.

Ima Okamura、Yoshihiko Yamazaki、Tokio Yamaguchi、Yasuo Komoda、Akira Nakajima：“抗胆红素单克隆抗体。III.非结合胆红素-IXα单克隆抗体的制备和特性”Biochimica et Biophysica Acta。

山崎 順彦: "抗ビリルビン単クロ-ン抗体を利用した血清ビリルビン測定の臨床的意義" 埼玉医科大学雑誌. 15. 325-338 (1988)

Yoshihiko Yamazaki：“使用抗胆红素单克隆抗体测量血清胆红素的临床意义”埼玉医科大学杂志 15. 325-338 (1988)。

Yuima OKAMURA: "Clinical significance of measurement of bilirubin in serum and urine using two kinds of anti-bilirubin monoclinal antibody" J. Saitama Med. School. 17. 235-251 (1990)

Yuima OKAMURA：“使用两种抗胆红素单克隆抗体测量血清和尿液中胆红素的临床意义”J. Saitama Med。

岡村 維摩: "エピト-プの異なる二種類の抗ビリルビン単クロ-ン抗体を利用した血清ビリルビン及び尿中ビリルビン測定の臨床的意義" 埼玉医科大学雑誌. 17. 235-251 (1990)

冈村今：“使用两种不同表位的抗胆红素单克隆抗体测量血清胆红素和尿胆红素的临床意义”，埼玉医科大学学报，17. 235-251 (1990)。