GENERATION OF HUMAN ANTI-TUMOR MONOCLONAL ANTIBODIES

人类抗肿瘤单克隆抗体的产生

• 批准号: 3460964
• 负责人: PETER BRAMS
• 金额: $ 10.95万
• 依托单位: IDEC PHARMACEUTICALS CORPORATION
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-24 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3460964
• 关键词: B lymphocyte; SCID mouse; antitumor antibody; genetic library; human genetic material tag; human tissue; hybridomas; molecular cloning; monoclonal antibody; neoplasm /cancer immunotherapy; passive immunization; technology /technique; tissue /cell culture; tumor antigens

The overall goal of this project is to produce human monoclonal antibodies against human tumor associated antigens (TAA). Two highly specific TAAs have been selected: a prostate specific antigen (PSA, 30 kD glycoprotein) and Carcinoembryonic antigen (CEA, 180 kD glycoprotein). Our approach is to utilize a combination of three methods previously independently used to generate human monoclonal antibodies: a) in vitro immunization of human spleen cells, b) immunization and expansion of human lymphocytes in SCID mice (hu-SCID model), and c) cloning and expression of human antibody genes by construction of M-13 combinatorial libraries. To achieve these goals, we have divided our project into 4 specific aims: I) In vitro immunization to generate B-cells primed against TAAs, II) immunization of SCID mice to further expand and select high affinity antibody producing human B-cell populations, III) construction of combinatorial libraries to clone and expand human antibody genes, and IV) antibody production and biological testing to obtain preclinical data on therapeutic human antibodies for passive immunotherapy. In preliminary experiments we could demonstrate that the combination of in vitro priming/in vivo boosting can yield significant titers of human antibodies against another human tumor associated antigen (ferritin). Furthermore, the feasibility of cloning human antibody genes from hu-SCID mice has recently been demonstrated (Nature (1992), 355:258). We believe that our novel combination approach can be extended to the generation of therapeutic antibodies against other diseases, beyond the currently targeted tumor associated antigens, and will become a general method for generating specific human MoAbs with high affinity.

本项目的总体目标是生产人源单克隆抗体 针对人肿瘤相关抗原（TAA）。两种高度特异性的TAA 已选定：前列腺特异性抗原（PSA，30 kD糖蛋白） 和癌胚抗原（CEA，180 kD糖蛋白）。 我们的方法是利用以前的三种方法的组合 独立地用于产生人单克隆抗体：a）体外 人脾细胞免疫，B）人的免疫和扩增 SCID小鼠中的淋巴细胞（hu-SCID模型），和c）克隆和表达 通过构建M-13组合文库获得人抗体基因。 为了实现这些目标，我们将我们的项目分为四个具体目标： I）体外免疫以产生针对TAA致敏的B细胞， 免疫SCID小鼠以进一步扩增和选择高亲和力 产生抗体的人B细胞群，III）构建 用于克隆和扩增人抗体基因的组合文库，和IV） 抗体生产和生物学测试，以获得临床前数据， 用于被动免疫治疗的治疗性人抗体。 在初步实验中，我们可以证明， 体外引发/体内加强可以产生显著的人免疫球蛋白的滴度。 抗另一种人肿瘤相关抗原（铁蛋白）的抗体。 进一步探讨了从hu-SCID中克隆人源抗体基因的可行性 最近已经证明了在小鼠中的应用（Nature（1992），355：258）。 我们相信，我们的新组合方法可以扩展到 产生针对其他疾病的治疗性抗体， 目前靶向肿瘤相关抗原，并将成为一个通用的 产生具有高亲和力的特异性人单克隆抗体的方法。