TGF-BETA 3 PRODUCTION IN EUCARYOTES AND PROCARYOTES

真核生物和原核生物中 TGF-β 3 的产生

• 批准号: 3506722
• 负责人: JOHN DOUGLAS HALEY
• 金额: $ 18.43万
• 依托单位: OSI PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-04-01 至 1992-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3506722
• 关键词: Escherichia coli; antineoplastics; biological models; biotechnology; cell differentiation; cell growth regulation; chimeric proteins; clone cells; complementary DNA; differentiation antigens; drug design /synthesis /production; drug screening /evaluation; gene expression; genetic promoter element; genetic translation; growth media; hematopoietic stem cells; intracellular transport; molecular cloning; monoclonal antibody; neoplastic cell; posttranscriptional RNA processing; protein biosynthesis; protein purification; protein transport; recombinant DNA; site directed mutagenesis; tissue /cell culture; transforming growth factors; transposon /insertion element

DESCRIPTION: (Adapted from the applicant's abstract) TGF-beta3 is a 25 kD polypeptide which exhibits potent biological effects on a variety of cell types. TGF-beta3 may be useful in cancer therapy, in wound healing indications and as an immunosuppressive agent. The goal of Phase I study was to express the TGF-beta3 cDNA. The investigators succeeded in establishing an initial expression system for TGF-beta3 and purified the corresponding protein to homogeneity. Furthermore, they prepared a series of both peptide polyclonal and monoclonal antibodies (MAbs) against different epitopes in TGF-beta3. Via their corporate collaboration with Pfizer, they now have a major pharmaceutical company interested in TGF-beta3 for multiple indications. However, it is apparent that commercial development is totally dependent upon establishing a cost effective production system. In Phase II study, they propose to increase the efficiency of expression of the recombinant protein produced in mammalian cells. Based on recent methodologies demonstrating successful expression of disulfide linked dimeric polypeptides in E. coli, they will also attempt to develop a procaryotic expression system to produce TGF-beta3 in E. coli.

描述：（改编自申请人的摘要）TGF-beta3 是一种 25 kD 对多种细胞表现出有效生物效应的多肽 类型。 TGF-β3 可能有助于癌症治疗和伤口愈合 适应症和作为免疫抑制剂。 一期研究的目标 目的是表达TGF-β3 cDNA。 调查人员成功 建立TGF-β3初始表达系统并纯化 相应的蛋白质同质性。 此外，他们还准备了一系列 肽多克隆抗体和单克隆抗体 (MAb) TGF-β3 中的不同表位。 通过他们的企业合作 辉瑞公司，他们现在有一家大型制药公司对 TGF-β3 适用于多种适应症。 然而，显而易见的是 商业发展完全取决于建立成本 有效的生产体系​​。 在第二阶段研究中，他们建议增加 产生的重组蛋白的表达效率 哺乳动物细胞。 基于最近证明成功的方法 在大肠杆菌中表达二硫键连接的二聚体多肽，它们将 还尝试开发原核表达系统来生产 大肠杆菌中的 TGF-β3。