ANTIBODIES TO ACTIVATED MUTANT RAS ONCOGENE PROTEINS

激活的突变 RAS 癌基因蛋白抗体

• 批准号: 3492194
• 负责人: FRED H REYNOLDS
• 金额: $ 5万
• 依托单位: OSI PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-05-01 至 1989-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3492194
• 关键词: affinity chromatography; antibody; antibody specificity; diagnosis design /evaluation; immunocytochemistry; immunologic techniques; immunological substance; immunoprecipitation; laboratory rabbit; monoclonal antibody; mutant; neoplasm /cancer immunodiagnosis; oncogenes; oncoproteins; protein purification; radiotracer; reagent /indicator; synthetic peptide; tissue /cell culture; tumor antigens; western blottings

Activated ras oncogenes have been identified in a significant percentage of cancers including leukemias and carcinomas of the colon, pancreas and lung. High affinity antibodies reactive with both normal and activated human p21ras are available. However, previous attempts to develop antibodies specific for mutant p21ras, by use of synthetic peptide immunogens, have not produced reagents of sufficient quality for clinical diagnostic assays. In order to obtain improved antibodies against activated ras proteins, recombinant p21ras protein, encoded by a K-ras gene with a mutation causing an aspartic acid for glycine substitution at residue 12, will be used to immunize rabbits. The desired antibodies will be purified by affinity chromatography on mutant p21ras, and antibodies reactive with normal ras proteins will be removed by absorption against wild-type p21ras. The antibodies will be assessed for binding to mutationally activated p21ras proteins, and will be used to develop both immunometric (antigen capture) and immunohistochemical assays for the mutant proteins. The ability to rapidly identify activated p21ras proteins in tumor biopsy specimens will greatly facilitate the progress of research on the clinical significance of ras oncogene activation, and should improve the specific diagnosis and prognostic evaluation of human cancer.

ras癌基因的激活已经在一个显著的 包括白血病和癌症在内的癌症的百分比 结肠胰腺和肺 高亲和力抗体与 正常的和活化的人p21 ras都是可用的。 然而，在这方面， 先前开发突变型p21 ras特异性抗体的尝试， 通过使用合成肽免疫原，还没有生产出试剂 具有足够的质量用于临床诊断测定。 为了 获得针对活化的ras蛋白的改进的抗体， 重组p21 ras蛋白，由具有突变的K-ras基因编码 导致在残基12处天冬氨酸被甘氨酸取代， 将用于免疫兔子。 所需的抗体将是 通过突变体p21 ras亲和层析纯化，和 与正常ras蛋白反应的抗体将被去除， 针对野生型p21 ras的吸收。 抗体会 评估与突变激活的p21 ras蛋白的结合，和 将用于开发免疫测定（抗原捕获）和 突变蛋白的免疫组织化学测定。 的能力 在肿瘤活检中快速鉴定活化的p21 ras蛋白 标本将大大促进研究的进展， ras癌基因激活的临床意义，并应 提高对人类肿瘤的特异性诊断和预后评估 癌