Production of purified plasmidial DNA and recombinant proteins, on a large scale, for use in vaccines and diagnostics

Abstract

The project aims to develop the production of plasmidial DNA and recombinant proteins to meet the demand of researchers and also to open perspectives for a new profile for the company. The project introduces significant technological innovation, given that the national market does not have access to such a product. In the first phase of development of the project studies were undertaken of the purification process of plasmidial DNA in the scale of 1 gram, meeting the criteria of regulatory requirements, which permitted the start of the planned clinical studies, presently in progress, applied to the treatment of epidermoid tumors in humans and bovine tuberculosis. From the information available, it is the first time that a company has mastered the technology of purification of plasmidial DNA on a large scale in Brazil. The competitive differential is immense and the costs of production are 15 to 20 times lower. Two commercial proposals are presently being analyzed, in order to subsidize the development and clinical study of two DNA vaccines in development with national researchers. Phase 1 of the project aimed to obtain the large scale production of DNA and its results proved to be totally viable and they are in harmony with the broader aims of the project. Namely, the production of a vaccine against tuberculosis, which was recently patented by FAPESP and the University of São Paulo, USP, and optimized by researchers from USP and the Federal University ofMinas Gerais, UFMG. The latter based itself on the incorporation of plasmidial DNA in biodegradable polymeric systems. In phase 2 of the project the aim is to obtain a stage of fermentation now on a semiindustrial scale (100 liters) and the production of micro- structured systems on a scale of 100 grams to 1 kilogram in order to enable clinical studies. The focus of this stage is the optimization of the production of plasmidial DNA and the application to other types of DNA in addition to the model worked on till then, which equates to the protein exposed to 65 KDa (hsp65) thermal shock and the development of the large scale production of non-living systems of transgenic infection based in biodegradable polymers and lipids on a pilot scale of cationic nano and microspheres and deprived of charges. (AU)