A GLUCOSE OXIDASE-BASED IMMUNOTOXIN PRODUCED IN YEAST

酵母产生的基于葡萄糖氧化酶的免疫毒素

• 批准号: 6142521
• 负责人: ROBERT V BLACKBURN
• 金额: $ 11万
• 依托单位: BIOSAVITA, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-08 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6142521
• 关键词: Saccharomyces cerevisiae; antineoplastics; athymic mouse; cell transformation; chimeric proteins; electroporation; expression cloning; glucose oxidase; immunological substance; immunotoxicity; monoclonal antibody; neoplastic cell; tissue /cell culture; tumor antigens; western blottings

This Phase I application offers a novel approach to the development of a recombinant immunotoxin consisting of a tumor antigen (EGP-2)-specific single chain antibody fused to the enzyme glucose-oxidase (GO). The goal of this proposal is to achieve high level expression of this immunotoxin in the yeast, saccharomyces cervisiae, and to determine the feasibility of its use as an anticancer agent. Preliminary evidence indicates that this GO-based immunotoxin exhibits specific binding to the EGP-2 antigen and enzymatic glucose oxidase activity (hydrogen peroxide production). Proposed in vitro studies will examine the effect of yeast produced immunotoxin on the survival of normal and EGP-2 positive human colon cancer cells. This revised Phase I application also includes limited in vivo studies to examine the anticancer effects of this immunotoxin against human colon cell tumors in a mouse xenograft model. Upon completion of the Phase I aims, we will establish proof of concept for the effectiveness of this novel GO-based immunotoxin produced in yeast Phase Il will further develop and optimize the expression and low cost production of other immunotoxins, and will demonstrate their therapeutic potential by in vitro and in vivo tests for the development of superior immunotherapy systems. PROPOSED COMMERCIAL APPLICATIONS: Immunotoxins and monoclonal antibody-based therapies are rapidly being developed by biopharmaceutical industries and the market potential for these drugs will be over $6 billion by year 2000. This Phase I proposal addresses two important issues for this market: development of novel immunotoxins as anti-cancer agents and optimization of low cost systems for the production of immunotoxins that will help reduce healthcare costs.

该I期申请提供了一种开发重组免疫毒素的新方法，该重组免疫毒素由融合至葡萄糖氧化酶（GO）的肿瘤抗原（EGP-2）特异性单链抗体组成。本提案的目的是实现这种免疫毒素在酵母菌中的高水平表达，并确定其作为抗癌剂的可行性。初步证据表明，这种基于GO的免疫毒素表现出与EGP-2抗原的特异性结合和酶促葡萄糖氧化酶活性（过氧化氢产生）。 拟议的体外研究将检查酵母产生的免疫毒素对正常和EGP-2阳性人结肠癌细胞存活的影响。该修订的I期申请还包括有限的体内研究，以检查该免疫毒素在小鼠异种移植模型中对人结肠细胞肿瘤的抗癌作用。在完成I期目标后，我们将为在酵母中产生的这种新型基于GO的免疫毒素的有效性建立概念证明。II期将进一步开发和优化其他免疫毒素的表达和低成本生产，并将通过体外和体内试验证明它们的治疗潜力，以开发上级免疫治疗系统。拟议的商业应用：生物制药工业正在迅速开发基于免疫毒素和单克隆抗体的疗法，到2000年，这些药物的市场潜力将超过60亿美元。 该第一阶段提案解决了该市场的两个重要问题：开发新型免疫毒素作为抗癌药物，以及优化用于生产免疫毒素的低成本系统，这将有助于降低医疗保健成本。