LUMINESCENCE DETECTION REAGENTS FOR GLYCOLIPIDS

糖脂发光检测试剂

• 批准号: 3489095
• 负责人: DAVID F SMITH
• 金额: $ 5万
• 依托单位: ELA TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-15 至 1990-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3489095
• 关键词: binding proteins; biomedical equipment development; biotin; diagnosis design /evaluation; glycolipids; glycosphingolipids; immunochemistry; lectin; luminescence; monoclonal antibody; neoplasm /cancer immunodiagnosis; reagent /indicator; thin layer chromatography; tumor antigens

Alteration in surface carbohydrates in glycolipids and glcoproteins of tumor cells relative to normal cells is a common feature of malignant diseases. Immunostaining procedures developed specifically for detecting glycolipid antigens with tumor specific antibodies represent important new techniques for immunodiagnostics and immunotherapeutics. The current methods for these diagnostic tests rely on reagents that are radioactive or that are enzyme-linked. In the studies proposed a new and general method will be developed for assaying glycolipids on solid supports that is more rapid and 3 to 6 orders of magnitude more sensitive than existing techniques and that does not utilize radioisotopes. This new technology exploits the remarkable sensitivity of the recombinant, bioluminescent protein, aequorin, and the high affinity interactions of streptavidin and biotin. The work described in this proposal has three specific aims. Fist, assays will be established using luminescence detection reagents (LDR's) for assaying Forssman glycolipid, a human tumor antigen, in microtiter plates. Second, this assay will be adapted to detecting glycolipids separated by thin layer chromatography. Third, an assay will be developed for detecting the sialyl-Le a antigen in complex mixtures using LDR in combination with the colorectalcarcinoma specific monoclonal antibody CA 19-9. The assays and reagents developed during this study will have important applications in both basic research and clinical laboratories involved in analysis of complex carbohydrates.

糖脂和糖蛋白表面糖的变化 肿瘤细胞相对于正常细胞是恶性肿瘤的共同特征， 疾病免疫染色程序专门用于检测 糖脂抗原与肿瘤特异性抗体代表重要的 免疫诊断和免疫治疗的新技术。 的 目前用于这些诊断测试的方法依赖于 放射性的或酶连接的。 在研究中提出了一种新的 并建立了测定固体糖脂的通用方法 支持的速度更快， 比现有技术敏感，并且不利用 放射性同位素 这项新技术利用了 的重组，生物发光蛋白，水母发光蛋白，和高 链霉亲和素和生物素的亲和相互作用。 中描述的工作 这项建议有三个具体目标。 首先，将建立分析 使用发光检测试剂（LDR's）测定福斯曼 糖脂，一种人类肿瘤抗原，在微量滴定板中。 二是这个 该测定法将适于检测通过薄层分离的糖脂 层析 第三，将开发一种用于检测 使用LDR与 结直肠癌特异性单克隆抗体CA 19-9。 的 本研究期间开发的测定和试剂将具有重要的 在基础研究和临床实验室中的应用 复杂碳水化合物的分析。