IMMUNOAFFINITY PURIFICATION WITH ANTI-ID ANTIBODIES

使用抗 ID 抗体进行免疫亲和纯化

• 批准号: 3492523
• 负责人: CARL Venton HAMBY
• 金额: $ 5万
• 依托单位: VIRO DYNAMICS CORPORATION
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-04-06 至 1990-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3492523
• 关键词: affinity chromatography; antiidiotype antibody; athymic mouse; computed axial tomography; diethylenetriaminepentaacetate; gel electrophoresis; hybridomas; melanoma; monoclonal antibody; neoplasm /cancer immunodiagnosis; neoplasm /cancer radiodiagnosis; protein purification; radiotracer; tissue /cell culture

One of the primary limitations in the use of radioimmunoscintigraphic techniques to detect tumors clinically has been the difficulty in achieving high tumor to normal tissue ratios of bound radioactivity. Part of this difficulty arises from the fact that a significant proportion of monoclonal antibodies (MoAb) are inactivated during radiolabeling procedures and such MoAb, when injected intravenously, localize non-specifically in normal tissues. This proposal addresses the problem through the development of an innovative, new method for effectively separating antibodies damaged during radiolabeling procedures from those retaining their original affinity and specificity. Preliminary studies indicate that anti-idiotypic (anti-Id) MoAb recognizing the combining sites of anti-melanoma MoAb can be prepared as affinity matrices to purify the anti-melanoma MoAb after radiolabeling procedures. In this study the immunoreactivity, specificity, affinity and tumor to tissue ratios of In-111-labeled anti-melanoma MoAb will be assayed before and after immunoaffinity is effective, it may be possible to increase the sensitivity of tumor detection by radioimmunoscintigraphy. Therefore we believe a ready commercial market exists among laboratories performing clinical or experimental radioimmunoscintigraphy techniques for kits containing anti-tumor MoAb ready for radiolabeling and prepackaged anti-Id MoAb affinity columns for purification after the MoAb have been labeled.

使用放射免疫闪烁显像的主要限制之一 临床检测肿瘤的技术一直是实现的难点 肿瘤与正常组织的结合放射性比率高。 这一部分 困难的产生是因为很大一部分单克隆抗体 抗体 (MoAb) 在放射性标记程序等过程中失活 MoAb，当静脉注射时，在正常情况下非特异性定位 组织。 该提案通过开发一个解决该问题的方法 有效分离过程中受损抗体的创新、新方法 保留其原始亲和力的放射性标记程序 特异性。 初步研究表明，抗独特型（anti-Id） 可以制备识别抗黑色素瘤MoAb结合位点的MoAb 作为亲和基质，在放射性标记后纯化抗黑色素瘤单克隆抗体 程序。 在这项研究中，免疫反应性、特异性、亲和力和 将测定 In-111 标记的抗黑色素瘤 MoAb 的肿瘤与组织比率 在免疫亲和力生效之前和之后，有可能 提高放射免疫闪烁扫描肿瘤检测的灵敏度。 因此，我们相信实验室之间存在现成的商业市场 执行临床或实验放射免疫闪烁扫描技术 含有抗肿瘤单克隆抗体的试剂盒，可用于放射性标记并预先包装 抗 Id MoAb 亲和柱，用于 MoAb 纯化后的纯化 标记。