SBIR-STTR Award

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

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.

Thesaurus Terms:
cost effectiveness, gene expression, method development, peptide chemical synthesis, protein purification analog, chimeric protein, drug discovery /isolation, elastin, phase change, protein folding, protein structure function CHO cell, Escherichia coli, combinatorial chemistry, liquid chromatography, mass spectrometry, microorganism culture