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Development of new purification tools for recombinant proteins

A recombinant protein purification platform based on cim(r) convective interaction media monolith technology will be developed to significantly decrease protein purification time and costs. The project will result in novel, technical, superior products and services and lower production costs.

PROTEIN LABORATORIES REHOVOT LTD (PLR) founded in August 2003 produces biologically active recombinant proteins (mainly different cytokines and their soluble receptors) in E. coli as well as expression plasmids and recombinant proteins upon request. The process starts from planning the optimised DNA (Deoxyribose Nucleic Acid) sequence, over-expressing it as insoluble protein (so called inclusion bodies), purifying the inclusion bodies and finally refolding it as a biologically active protein and purifying to over 98% homogeneity by ion-exchange and/or gel filtration chromatography. An alternative approach is to express the protein as a soluble protein in cytosol of bacteria. In such case the protein is already refolded but its purification is more complicated as it requires separation from other cytosolic proteins and fast purification to avoid proteolysis. The bottlenecks in this process are the purification steps, especially the time consuming gel filtration chromatography. As the purification represents 70-80% of the protein production costs any improvement would result in a considerable cost reduction and consequently, lower price of the product for the market. BIA SEPARATIONS D.O.O. founded in 1998 develops and produces CIM(R) Monolithic Columns. These revolutionary chromatographic supports are purpose designed for large biomolecule (pDNA (Plasmid DNA), virus, protein) purification and benefit convective mass transport which results in much faster separations (biomolecules maintain their activity), high volumetric throughput, and low back pressure. * The first goal of the project is to develop a new platform purification process which will replace time consuming gel filtration chromatography and provide PLR with an efficient tool for purification of proteins. Several approaches incorporating tangential flow filtration, ultra filtration, membrane adsorbers and CIM(R) monolithic columns will be tested and the best set-up of purification steps will be optimised. A new purification platform will be compared to the present protocols for different recombinant proteins produced by PLR and subsequently used for development of new recombinant proteins. Due to optimised purification process production costs for PLR it will be lower and it will speed the introduction of their products to the market. Consequently their customers would benefit with lower prices. * The second goal is to develop and test new purpose designed CIM(R) monolithic columns for purification of recombinant proteins. These new products will enable BIA SEPARATIONS to attack the recombinant protein purification market more aggressively on a+ global scale. * The third goal is to test novel Bio-Monolith columns, based on CIM(R) Monolith technology and marketed by Agilent Technologies for fast in-process control to replace time consuming SDS-PAGE (Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis) methods. New applications will help BIA SEPARATIONS to strengthen the position of analytical scale monoliths on the market. * The fourth goal is to develop and maintain co-operation between PLR and BIA SEPARATIONS. Partners are compatible with PLR producing recombinant proteins and BIA SEPARATIONS producing novel purification tools for these proteins. PLR will have an access to new state-of-the-art purification tools while BIA SEPARATIONS will be able to test their products on different proteins. The project is intended to start at the beginning of 2010 and lasts for 3 years, with BIA SEPARATIONS as project co-coordinator. R&D groups of both companies will be involved in the project. The PLR group will focus on protein production and optimisation of purification process, while the BIA SEPARATIONS group will focus on the development of purpose designed columns and their integration in a new more efficient platform purification process as well as in in-process control method development.
Acronym: 
RecPur
Project ID: 
5 347
Start date: 
01-01-2010
Project Duration: 
36months
Project costs: 
630 000.00€
Technological Area: 
Cellular and Molecular Biology
Market Area: 
Pharmaceuticals/fine chemicals

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