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）：本应用的长期目标是开发一种新型的蛋白质纯化技术，这将消除对色谱的需求。这项技术是Deltaphase TM技术，具有更大的选择性，更易于操作，并且比色谱法便宜得多。简而言之，三倍酶技术将改变蛋白质纯化的经济学。该技术的影响将在两个与健康有关的领域中： 新型药物的开发。由于通过功能基因组学确定了独特的药物靶标，因此需要表达和纯化以进行评估（目标验证）和筛选（高通量筛选和X射线晶体学）。相比技术将迅速扩展靶蛋白的池1）由于技术的显着溶解度增强特性和2），可以通过简单的一个步骤纯化方法来增加纯化的渗透率，从而在选择的表达系统（例如大肠杆菌）中启用蛋白的首次表达（例如，大肠杆菌）。 生物治疗药的较低成本。创新的生物制药公司使用Deltaphase TM技术将估计将纯化成本降低10倍。鉴于托管护理的定价压力，生物制药公司在降低商品成本时将具有更大的定价灵活性。 此外，由于制造成本如此之高，因此生物基市场尚未完全发展，并且通用和原始产品之间的价格差异尚未保证通用替代品的发展。 Deltaphase TM技术降低纯化成本的能力可能是生物基市场发展的重要催化剂。 该技术依赖于使用与感兴趣蛋白质融合的弹性蛋白样多肽（ELP）。这些标签使用重组DNA技术融合。 该项目的具体目的包括1。设计和合成，用于优化纯化2。ELP的表达和表征和3。四个药物相关的重组ELP融合蛋白的表达和纯化。该项目的里程碑包括：1。通过SDS-PAGE测量，将四种蛋白质中的两种纯化至少90％纯度。 2。对于两种纯化的蛋白质，一种蛋白质的活性将与该蛋白质相等的活性。 3。证明其中一种纯化蛋白的纯化成本（时间和材料）至少下降了两倍。