The expression of human catalytic proteins in micro algae on a commercial scale

人类催化蛋白在微藻中的商业规模表达

• 批准号: 7235234
• 负责人: Richard Thomas Sayre
• 金额: $ 55.97万
• 依托单位: U.S. ARMY MEDICAL RESEARCH INST CHEM DEF
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7235234
• 关键词: algae; human; plants; proteins

Bioscavenging of organophosphate (OP) by human proteins is emerging as a promising medical intervention for prophylaxis and post-exposure treatment against chemical warfare nerve agents. The best-.studied bioscavengers (BSCs) to date, meeting considerable success in pre-clinical research, are human cholinesterases (ChEs). However, ChEs, which are highly efficient in binding and sequestering OPs, are also inactivated by the toxins and therefore administration of large amounts of protein is necessary for full protection, raising the question of the practicality of this approach. The development of improved biocatalysts (BCT) (paraoxonase 1) that can catalytically degrade OPs may address this concern. The proposed effort offers a novel means to biomanufacture recombinant BSCs and BCTs based on the human proteins, butyrylcholinesterase and paraoxonase 1. In collaboration with the other projects participating in the Center, the capacity of these proteins to sequester or hydrolyze OPs will be improved by subjecting their genes to either random in vitro evolution or rational mutagenesis. To that end, we will undertake high-throughput screening of mutant enzyme libraries, in particular to improve the stability and drug-like properties of paraoxonase I. In addition, post-translational glycosylation systems or protein PEGylation procedures will be developed to increase the circulating lifetimes and to eliminate potential antigenicity of proteins produced in non-human, recombinant organisms. The OSU team (Project 6) will utilize the transgenic microalgae, Chlamydomonas reinhardtii, to produce pilot-plant scale quantities of these products for direct recovery from the culture media. In addition, they will optimize this production system for scale-up to commercial production capacity. The primary significance of this project is that it will develop pilot-plant systems for the production of enhanced, second-generation, improved ChE-based BSC and BCT products suitable for human clinical trials.

人类蛋白质对有机磷（OP）的生物清除正在成为一种有前途的医疗干预措施 用于化学战神经毒剂的预防和暴露后治疗。研究得最好的 迄今为止，生物清除剂（BSC）在临床前研究中取得了相当大的成功，都是人类的 胆碱酯酶（ChE）。然而，ChE 在结合和隔离 OP 方面非常高效， 被毒素灭活，因此需要施用大量蛋白质才能充分发挥作用。 保护，提出了这种方法的实用性问题。改进生物催化剂的开发 (BCT)（对氧磷酶 1）可以催化降解 OP，可能会解决这个问题。 拟议的工作提供了一种基于生物制造重组 BSC 和 BCT 的新方法 人类蛋白质、丁酰胆碱酯酶和对氧磷酶 1. 与其他项目合作 参与该中心后，这些蛋白质螯合或水解 OP 的能力将通过以下方式得到提高： 让他们的基因经历随机的体外进化或合理的诱变。为此，我们将 对突变酶库进行高通量筛选，特别是提高稳定性和 对氧磷酶 I 的类似药物特性。此外，翻译后糖基化系统或蛋白质 将开发聚乙二醇化程序以增加循环寿命并消除潜在的 非人类重组生物体中产生的蛋白质的抗原性。 OSU 团队（项目 6）将 利用转基因微藻莱茵衣藻生产这些中试规模的微藻 用于直接从培养基中回收的产品。此外，他们还将优化该生产系统 扩大到商业生产能力。该项目的首要意义在于，它将发展 用于生产增强型、第二代、改进的基于 ChE 的 BSC 和 BCT 的中试工厂系统 适合人体临床试验的产品。