Low Cost Production of Recombinant Proteins

描述（由申请人提供）：有越来越多的蛋白质药物在管道中，与合成的小分子药物相比，其生产成本非常高。此外，一些蛋白质药物的专利将在未来几年内到期，这将为这些药物的仿制药打开市场，如果它们能够以更低的成本生产并证明生物等效性的话。I期研究结果清楚地表明，deltphase系统在蛋白质重组生产方面具有强大的商业价值，其中一些蛋白质难以作为可溶性蛋白高水平表达。由于重组蛋白药物的重要性日益增加，以及对低成本表达和易于纯化蛋白质的明确需求，II期研究的目标将是推进重组蛋白的表达和纯化技术，使其在未来三年内具有高表达和高纯度水平的生物仿制药资格。使Phase Bioscience能够实现这一目标的技术是deltaPhaseTM系统，这是一种新型平台技术，用于高水平表达和简单的非色谱纯化，使用一类称为弹性蛋白样多肽（ELPs）的多肽标签。该技术的基本基础是发现融合到ELP的蛋白质和肽保留了ELP的环境触发（温度，盐）可溶-不溶性相变行为，并且这种可逆相变可以在细胞裂解液中进行，通过几个相变循环将ELP融合物与其他细胞蛋白分离到均匀性。该技术的影响将是为蛋白质行业提供生产蛋白质的能力，特别是生物仿制蛋白质，通过从纯化过程中去除色谱，以显著降低成本。这将使这些药物与小分子药物相比更具竞争力，并降低患者的药物成本。该项目的具体目标包括：1。在大肠杆菌和CHO中表达6种具有药学意义的生物仿制候选重组融合蛋白2. 用LC-MS将蛋白纯化至至少90%的纯度；和3。与商业生产的蛋白质相比，证明它们的生物活性。

DESCRIPTION (provided by applicant): There are an increasing number of protein drugs in the pipeline, whose cost of production is very high compared to synthetic, small molecule drugs. Further, a number of protein drugs are coming off patent in the next few years, which will create an opening for a market for generic version of these drugs, if they can be produced at significantly lower cost and demonstrate bioequivalence. The Phase I results clearly indicated that a powerful and commercially important application of the deltaPhase System is in the recombinant production of proteins, some of which are difficult to express at high levels as soluble proteins. Motivated by the increasing importance of recombinant protein drugs, and the clear need for low-cost expression and easy purification of proteins, the goal of the Phase II research will be to advance the technology for the expression and purification of recombinant proteins that will be eligible for biogenerics within the next three years, with high expression and at high purity levels. The enabling technology that will allow Phase Bioscience to achieve this objective is the deltaPhaseTM System, a novel platform technology for high-level expression and simple, non-chromatographic purification using a class of polypeptide tags, termed elastin-like polypeptides (ELPs). The fundamental basis of this technology is the discovery that proteins and peptides fused to an ELP retain the environmentally triggered (temperature, salt) soluble-insoluble phase transition behavior of ELPs, and that this reversible phase transition can be carried out in cell lysate to separate ELP fusions from other cellular proteins to homogeneity by a few phase transition cycles. The impact of the technology will be to provide the protein industry with the ability to produce proteins, especially biogeneric proteins, at significantly lower cost by removing chromatography from the purification process. This will make these drugs more competitive with small molecule drugs, and reduce the cost of drugs to patients. The Specific Aims of the project include: 1. Express six pharmaceutically-relevant, biogeneric candidate recombinant fusion proteins in E. coli and CHO; 2. Purify the proteins to at least 90% purity, as measured by LC-MS; and 3. Demonstrate their bioactivity compared to the commercially produced proteins.