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Stability and activity of the anti-leukemic L-asparaginase in the presence of different ionic liquids and polymers

Grant number: 16/09606-7
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2016
Effective date (End): May 31, 2017
Field of knowledge:Engineering - Chemical Engineering
Principal Investigator:Jorge Fernando Brandão Pereira
Grantee:Thainá de Souza Pecorari
Home Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
Associated research grant:14/16424-7 - Optimization and scale-up of liquid-liquid extraction process with ionic liquids (ILs) as a sustainable tool for the separation of the anti-leukemia biopharmaceutical L-asparaginase (ASPase), AP.JP

Abstract

The search for new pharmaceuticals for the treatment of hematologic patients have induced an increment on the number of studies with the aim of the production and purification of several biopharmaceuticals. Among them, the L-asparaginase (ASNase), a tetrameric enzyme used in chemoterapic sessions, have enlarged as one of the most promising for the Childhood Acute Lymphoblastic Leukemia Treatment (LLA). However, its purification is still one of the limiting steps, in particular, when the enzyme is produced through fermentation processes, since it is normally produced in very low concentrations. Their initial purification is based in traditional liquid-liquid extraction processes, which can purify and concentrate the enzyme. Among these, the possibility of using aqueous two-phase systems (ATPS) as alternative techniques of liquid-liquid extraction, have generated the submission of the Young Researcher project, which is intended to purify the ASNase from fermented media using ATPS composed of ILs and polymers. Thus, to validate the use of these ATPS is recquired to assess whether these compounds do not destabilize the ASNase, and maintain its stability and enzymatic activity. Considering that purpose, the objective of this IC project is to evaluate what are the ILs and Polymers (different Polyethylene Glycols (PEGs)) that does not destabilize or inactivate the ASNase. Thus, the stability and activity of commercial ASPase to different concentrations of ILs and PEGs, different temperatures and pH values will be assessed. This project intends to define a series of compounds (PEGs and ILs) which do not adversely affect ASNase and can be subsequently used in purification of the enzyme from the fermented medium from recombinant P. pastoris. (AU)