Therapeutic l-asp for LLA purification employing alternative bioprocess

Abstract

There is a strong global trend to seek alternative routes for the production of active pharmaceutical ingredients obtained by biotechnological processes. However, in both research and in the pharmaceutical industrial production, Brazil has not only a weak expression in the world's scenario but also is losing its international biopharmaceuticals suppliers that are no longer interested in the market and started discontinuing the production of several of these, especially those related to the onco-hematology treatment. In this context, this project proposes the pre-purification of L-asparaginase, biopharmaceutical widely used to treat leucemia as Acute lymphoid leucemia (ALL) that mainly affects children 3-5 years old and adolescents, increasing its survival from 20% to 90%. This purification will be done, by precipitation with ammonium sulfate and ethanol integrated with the enzyme extraction by aqueous two-phase systems (PEG/polyacrylate system and micellar system using Titon X-114 and buffer). The pre-purification tests will be preceded by stability studies of commercial biomolecule and biosynthesized in the presence of the components used for the precipitation and extraction systems. The use of such systems is broad and can be applied at different stages of the enzyme production process, as in steps: clarification, concentration and purification. (AU)