Detection of Novel Amyloid-Associated Protein by Monoclonal Antibody and its Gene Analysis

新型淀粉样蛋白相关蛋白的单克隆抗体检测及其基因分析

基本信息

批准号：
01570363
负责人：
IMAI Kohzoh
金额：
$ 1.41万
依托单位：
Sapporo Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
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

项目摘要

We have found that monoclonal antibody AM34 (IgGl) recognizes a novel antigen which exists in amyloid deposits in various organs of the rheumatoid arthritis.In addition, anti-beta-protein monoclonal antibodies (TB1, TB2, TB3, TB4 and TB5) were generated from the fusion of mouse myeloma X63-Ag8.653 cells with splenocytes obtained from a BALB/c mouse that had been immunized with a synthetic peptide of beta-protein consisting of 24 amino acids. All these antibodies bound specifically to the amyloid changes of senile plaque and amyloid angiopathy of the brain tissues of patients with Alzheimer's disease (AD) or with senile dementia Alzheimer type (SDAT) , but not at all to protein in the normal brain or other tissues by the immunoperoxidase method. Formic acid pretreatment of brain tissue enhanced this reactivity. Positive reactions were clearly visible even in small deposits of plaque, senile plaque crown amyloid and amyloid deposits of capillary vessels under this condition.Western blot analysis revealed that monoclonal antibodies TB2 and TB3 reacted mainly with 116 kD band when extract of SDAT brain was used. They failed to react, however, with 116kD band when extract of normal brain derived from patients with other diseases was used. Two dimensional electrophoresis revealed that the 116 kD band migrated at around pl8.5. MoAbs TB1, TB2 and TB4 strongly reacted with the 1st part (7 amino acids near to N-terminus) of the synthetic peptide (24 amino acids) of the beta-protein, which consists of hydrophilic amino acids.

我们已经发现，单克隆抗体AM34（IgGL）识别出一种新型抗原，该抗原存在于类风湿关节炎的各种器官中的淀粉样蛋白沉积物中。此外，抗β-蛋白质单克隆单克隆抗体（TB1，TB2，TB2，TB3，TB4和TB5）是FBETA单克隆抗体（TB3，TB4和TB5）是FORMAS的FORMOS，是FORMOS的FISEL HORMO;用从BALB/C小鼠中获得的脾细胞，该BALB/C小鼠已用由24个氨基酸组成的β-蛋白质的合成肽免疫。所有这些抗体专门与阿尔茨海默氏病（AD）患者的老年斑块和淀粉样蛋白血管病的淀粉样蛋白变化或淀粉样蛋白症状的淀粉样蛋白（AD）或老年痴呆症阿尔茨海默氏症类型（SDAT），但根本没有对正常脑或其他组织中的蛋白质的蛋白质或其他组织。甲酸对脑组织的预处理增强了这种反应性。即使在这种情况下的小斑块，老年斑块冠淀粉样蛋白和淀粉样蛋白沉积物中，也可以清楚地看到阳性反应。Western印迹分析表明，当使用SDAT脑提取提取物时，单克隆抗体TB2和TB3主要与116 kd的单克隆抗体TB2和TB3反应。但是，当使用了从患有其他疾病的患者衍生出的正常脑提取的提取物时，他们无法与116kD的伴侣做出反应。二维电泳表明，116 kD带迁移在PL8.5左右。 MoAbs TB1，TB2和TB4与β-蛋白质的合成肽（24个氨基酸）的第一部分（接近N末端的7个氨基酸）反应，由亲水氨基酸组成。