PURIFICATION OF TUMOR ANTIGENS OF DEFINED SPECIFICITIES

纯化具有确定特异性的肿瘤抗原

• 批准号: 3168981
• 负责人: Rishab K. Gupta
• 金额: $ 14.42万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-04-01 至 1990-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3168981
• 关键词: antibody formation; chromatography; diagnosis quality /standard; enzyme linked immunosorbent assay; hybridomas; laboratory mouse; melanoma; monoclonal antibody; neoplasm /cancer immunodiagnosis; neoplasm /cancer immunology; prognosis; radioimmunoassay; tissue /cell culture; tumor antigens

The overall objective of this research is to purify tumor antigens of defined specificities from spent culture medium of melanoma cell lines and develop sensitive immunological tests, e.g., radioimmunoassay (RIA) and enzyme immunoassay (EIA), for detection and quantification of such antigens in the serum and other body fluids of cancer patients. Two antigens have been detected in the spent culture medium of a melanoma cell line that are immunogenic in the host. One antigen is of fetal origin and has been termed as fetal antigen (FA). FA is a glycoprotein of 60 to 70 kilodaltons. The other antigen is a lipoprotein of 180 to 190 kilodaltons, and has been observed to be preferentially expressed by melanoma cells and rarely by other tumor cells. Procedures have been developed to detect the presence of melanoma-associated antigen (MAA) circulation in human sera, both in free form and in association with immune complexes. The MAA has been further characterized physicochemically. It has a beta-electophoretic mobility. Upon delipidation of MAA with 2-chloroethanol and further purification by Sephadex LH 20 chromatography, it was observed that the protein portion is comprised of two polypeptides. These polypeptides are 17.5 and 26 kilodaltons as determined by SDS-PAGE under reducing condition. In addition, affinity procedures coupled with immunoprecipitation with allogeneic antibody have been developed to identify various antigenic components of immune complexes circulating in melanoma patients. In one instance, at least five different polypeptide chains were recognized. Of these, two subunits, 92 and 41 kilodaltons were predominant. These polypeptides are quite different, at least on the basis of molecular weight, from MAA. (2)

本研究的总体目标是纯化肿瘤抗原 黑色素瘤细胞系用过的培养基的明确特异性 开发敏感的免疫学测试，例如放射免疫测定（RIA）和 酶免疫分析 (EIA)，用于检测和定量此类抗原 癌症患者的血清和其他体液中。 在黑色素瘤的废培养基中检测到两种抗原 在宿主中具有免疫原性的细胞系。 一种抗原源自胎儿 并被称为胎儿抗原（FA）。 FA是一种糖蛋白，分子量为60至70 千道尔顿。 另一种抗原是180至190道尔顿的脂蛋白， 并已观察到优先由黑色素瘤细胞表达 很少由其他肿瘤细胞引起。 已制定程序来检测 人类血清中存在黑色素瘤相关抗原（MAA）循环， 既有游离形式，也有与免疫复合物结合的形式。 MAA 有 进一步进行了物理化学表征。 它具有β-电泳 流动性。 MAA 用 2-氯乙醇脱脂并进一步 通过 Sephadex LH 20 色谱纯化，观察到 蛋白质部分由两条多肽组成。 这些多肽是 在还原条件下通过 SDS-PAGE 测定为 17.5 和 26 千道尔顿 健康）状况。 此外，亲和程序与 同种异体抗体免疫沉淀法已被开发用于 识别循环中免疫复合物的各种抗原成分 黑色素瘤患者。 在一种情况下，至少五种不同的多肽 连锁店得到了认可。 其中，两个亚基，92 和 41 千道尔顿 占主导地位。 这些多肽是相当不同的，至少在基础上是这样的 分子量，来自MAA。 (2)