SBIR Phase II: Multivariate Analysis of Heterologous Protein Expression

这项SBIR II期研究开发了改进在国外宿主中生产的重组蛋白产品的制造方法。具有成本效益的蛋白质生产通常利用非常适合蛋白质工程和大规模生产的生物。建立一个合适的蛋白质生产系统通常是一个耗时的、基于试错的过程，并且可能是蛋白质商业化的一个重大障碍。在发现生产系统的情况下，由于所需的时间和成本以及我们目前对关键参数的有限理解，它们通常远未优化。在第一阶段，评估了几个基因设计变量对大肠杆菌蛋白表达的重要性，大肠杆菌是一种常用的生产生物。数据表明，基因优化的新手段是意想不到的，从传统智慧。在第二阶段，将探索第一阶段提出的相关基因设计变量，以建立一个完善的基因设计与大肠杆菌和其他有用生产生物中蛋白质表达关系的模型。这项研究的更广泛的影响是改进了重组蛋白产品的制造技术。包括工业酶、诊断酶和蛋白质药物在内的蛋白质产品目前构成了一个价值400亿美元且快速增长的全球市场。该项目开发的工具将大大提高速度，降低成本，并消除现代蛋白质制造的不确定性，这些不确定性极大地限制了这一市场。提高产量还将加速对具有治疗或其他销售潜力的蛋白质的研究，扩大候选蛋白质的商业化领域。

This SBIR Phase II research develops methods to improve the manufacture of recombinant protein products produced in foreign hosts. Cost-effective production of proteins generally utilizes organisms that are well-suited for protein engineering and large-scale production. Establishing a suitable production system for a protein is often a time-consuming, trial-and-error-based process and can be a significant barrier for the commercialization of a protein. In cases where production systems are found, they are often far from optimized due to the time and cost required as well as our current limited understanding of the critical parameters. In Phase I several gene design variables were assessed for their importance to protein expression in the bacterium Escherichia coli, a commonly used production organism. Data suggested novel means for gene optimization that were unexpected from conventional wisdom. In Phase II relevant gene design variables suggested by Phase I will be explored toward development of a refined model of the relationship of gene design to protein expression in E. coli as well as in other useful production organisms.The broader impacts of this research are improved manufacturing techniques for recombinant protein based products. Protein products constitute a currently $40 billion and rapidly growing world-wide market including industrial enzymes, diagnostic enzymes and protein pharmaceuticals. The tools developed from this project will drastically improve the speed, reduce the cost, and remove the uncertainties of modern protein manufacturing, which significantly limit this market. Improved production will also accelerate the study of proteins with therapeutic or otherwise marketable potential, expanding the field of candidate proteins for commercialization.