Biopolymers for Relevant Recombinant Protein Puification

用于相关重组蛋白纯化的生物聚合物

• 批准号: 6785800
• 负责人: SUE DAGHER
• 金额: $ 10万
• 依托单位: PHASEBIO PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6785800
• 关键词: Escherichia coli; SDS polyacrylamide gel electrophoresis; affinity chromatography; androgen receptor; biotechnology; chimeric proteins; corticosteroid receptors; cost effectiveness; elastin; estrogen receptors; genetic manipulation; nucleic acid chemical synthesis; peptides; peroxisome proliferator activated receptor; polymers; protein purification; recombinant DNA; recombinant proteins; technology /technique development; transfection /expression vector

DESCRIPTION (provided by applicant): The long-term objective of this application is to develop a novel protein purification technology, which will eliminate the need for chromatography. This technology, the deltaPhase TM Technology, has greater selectivity, is simpler to operate and is much less expensive than chromatography. In short, the deltaPhase Technology will change the economics of protein purification. The impact of the technology will be in two health related areas: Development of Novel Medicines. As unique pharmaceutical targets are identified through functional genomics, they will need to be expressed and purified for evaluation (target validation) and screening (high-throughput screening and X-ray crystallography). The PhaseBio technology will rapidly expand the pool of target proteins by 1) enabling first-time expression of proteins in expression systems of choice (e.g., E. coli) due to the significant solubility enhancement properties of the technology and 2) increasing the throughput of purification by the simple, one step purification method which can be readily automated. Lower Cost of Bio-therapeutics. The use of the deltaPhase TM technology by innovative biopharmaceutical companies will reduce the purification costs by an estimated 10-fold. Given the pricing pressures by managed care, biopharmaceutical companies will have more pricing flexibility when the cost of goods is reduced. In addition, the biogeneric market has not fully developed because the cost of manufacturing has been so high, and the price differential between the generic and the original has not warranted development of the generic substitute. The ability of the deltaPhase TM technology to reduce purification costs could be an important catalyst in the development of a biogeneric market. The technology relies on the use of an Elastin-Like Polypeptide (ELP) fused to the protein of interest. These tags are fused using recombinant DNA techniques. The Specific Aims of the project include 1. Design and Synthesis of deltaPhase TM ELP tags for Optimized Purification 2. Expression and Characterization of ELPs and 3. Expression and Purification of Four Pharmaceutically Relevant Recombinant ELP Fusion Proteins. The Milestones for the project include: 1. Purify two of the four proteins to at least 90 percent purity, as measured by SDS-PAGE. 2. For the two purified proteins, one protein will have equal activity to that protein purified by conventional means. 3. Demonstrate at least a two-fold decrease in purification costs (time and material) for one of the purified proteins.

描述（由申请人提供）：本申请的长期目标是开发一种新型蛋白质纯化技术，该技术将消除对层析的需要。这项技术，即 deltaPhase TM 技术，比色谱法具有更高的选择性、操作更简单且成本低得多。简而言之，deltaPhase 技术将改变蛋白质纯化的经济性。该技术的影响将体现在两个与健康相关的领域： 新药开发。由于通过功能基因组学鉴定了独特的药物靶标，因此需要对其进行表达和纯化以进行评估（靶标验证）和筛选（高通量筛选和 X 射线晶体学）。 PhaseBio 技术将通过以下方式快速扩展目标蛋白库：1) 由于该技术具有显着的溶解度增强特性，使得蛋白质能够在所选表达系统（例如大肠杆菌）中首次表达；2) 提高蛋白质的通量通过简单的、易于自动化的一步纯化方法进行纯化。 生物治疗的成本较低。创新型生物制药公司使用 deltaPhase TM 技术预计可将纯化成本降低 10 倍。考虑到管理式医疗的定价压力，当商品成本降低时，生物制药公司将拥有更大的定价灵活性。 此外，生物仿制药市场尚未充分发展，因为制造成本如此之高，仿制药与原研药之间的价格差异不足以保证仿制药替代品的发展。 deltaPhase TM 技术降低纯化成本的能力可能成为生物仿制药市场发展的重要催化剂。 该技术依赖于使用与目标蛋白质融合的类弹性蛋白多肽（ELP）。这些标签通过重组 DNA 技术融合。 该项目的具体目标包括 1. 设计和合成用于优化纯化的 deltaPhase TM ELP 标签 2. ELP 的表达和表征以及 3. 四种药学相关重组 ELP 融合蛋白的表达和纯化。该项目的里程碑包括： 1. 根据 SDS-PAGE 测量，将四种蛋白质中的两种纯化至至少 90% 的纯度。 2. 对于两种纯化的蛋白质，其中一种蛋白质与通过常规方法纯化的蛋白质具有相同的活性。 3. 证明其中一种纯化蛋白质的纯化成本（时间和材料）至少降低两倍。