Biosynthesis and Downstream Recovery of Therapeutic Proteinsfrom the Milk of Transgenic Animals

转基因动物奶中治疗性蛋白质的生物合成和下游回收

• 批准号: 9011098
• 负责人: William Velander
• 金额: $ 56万
• 依托单位: Virginia Polytechnic Institute and State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-15 至 1995-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9011098&HistoricalAwards=false
• 关键词: Biosynthesis; Downstream; Recovery; Therapeutic; Proteinsfrom

Therapeutic proteins are an example of high-value proteins whose biosynthesis and downstream recovery has been limited by low productivity, extreme contamination, FDA requirements and difficulties due to improper modification or deleterious altering of the protein structure by the genetically engineered production system such as tissue culture. These problems have led to an undesirable mode of treatment available to people such as hemophiliacs due to cost and limited availability of the replacement protein. A detailed understanding of the structure and activity of the protein produced by the recombinant system facilitates the engineering of recovery methods for appropriate forms of the protein of interest as well as having genetic value for the recovery of other proteins using that same expression system. Transgenic mice which express human Protein C that is biologically active have been a good model for the production of a therapeutic protein by using a mammal as a bioreactor. Protein C is an anti-clotting protein which has potential for the treatment of clotting problems associated with septic shock, heart attack, and hip replacement surgery. The distribution of Protein C products is being evaluated based upon proteolytic and biosynthetic modifications characteristic of mammary tissue expression. Proteolysis seems to effect the integrity of the Protein C produced in murine milk. Pig milk doped with Protein C has also shown this same problem. Transgenic pigs containing the same genetic information as successfully used in mice are being evaluated as a large scale bioreactor with the potential of grams of heterologous protein per day. An immunoaffinity procedure for isolation of Protein C from murine and pig milk is being developed. The future use of transgenic animal bioreactors depends upon the knowledge of the biochemical changes that are affected by the mammary tissue and milk environments.

治疗性蛋白质是高价值的 蛋白质的生物合成和下游回收， 受到低生产率、极端污染 FDA的要求和困难，由于不当 蛋白质的修饰或有害改变 基因工程生产系统 例如组织培养。 这些问题导致了 不受欢迎的治疗方式， 血友病患者由于成本和有限的可用性， 替代蛋白 详细了解 蛋白质的结构和活性 重组系统促进了 适当形式的蛋白质的回收方法 兴趣以及具有遗传价值的恢复 使用相同的表达系统。 表达人蛋白C的转基因小鼠， 生物活性一直是一个很好的模型， 通过使用哺乳动物作为 生物反应器。 蛋白C是一种抗凝血蛋白 它有治疗凝血的潜力 与败血性休克、心脏病发作和 髋关节置换手术 蛋白C的分布 产品是基于蛋白水解和 乳腺癌的生物合成特征 组织表达 蛋白质水解似乎会影响 小鼠乳汁中产生的蛋白C的完整性。 猪 掺入蛋白C的牛奶也表现出同样的情况 问题. 含有相同基因的转基因猪 在小鼠中成功使用的信息正在被 评价为具有潜在的大规模生物反应器 的异源蛋白质。 一个 用于分离蛋白C免疫亲和方法 正在开发鼠奶和猪奶。 未来使用 转基因动物生物反应器取决于 生物化学变化的知识，受影响的 乳腺组织和乳汁环境。