LUMINESCENCE DETECTION REAGENTS FOR FOOD PATHOGENS

红豆杉糖脂和糖蛋白表面碳水化合物的变化 肿瘤细胞相对于正常细胞是恶性肿瘤的共同特征 疾病。免疫染色程序专门为检测 糖脂抗原和肿瘤特异性抗体是重要的 免疫诊断和免疫治疗的新技术。这个 目前用于这些诊断测试的方法依赖于 放射性的或与酶相关的。在研究中提出了一种新的 并将开发出测定固体糖脂的通用方法 支持速度更快，多3到6个数量级 比现有技术更敏感，而且没有利用 放射性同位素。这项新技术利用了显著的敏感性 重组生物发光蛋白Aequorin和HIGH 链霉亲和素与生物素的亲和作用。中描述的工作 这项提议有三个具体目标。首先，将建立检测方法 用发光检测试剂(LDR‘s)测定Forssman 糖脂，一种人类肿瘤抗原，在微滴定板中。第二，这个 检测薄层分离糖脂的方法将得到改进 层析法。第三，将开发一种检测方法来检测 LDR联合应用于复杂混合物中唾液酸乐抗原的研究 结直肠癌特异性单抗CA19-9。这个 在这项研究中开发的分析和试剂将具有重要的 在基础研究和临床实验室中的应用 在复杂碳水化合物的分析中。

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.