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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Controlling the l-asparaginase extraction and purification by the appropriate selection of polymer/salt-based aqueous biphasic systems

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Author(s):
Magri, Agnes [1] ; Pimenta, Marcela V. [2] ; Santos, Joao H. P. M. [2, 3] ; Coutinho, Joao A. P. [3] ; Ventura, Sonia P. M. [3] ; Monteiro, Gisele [2] ; Rangel-Yagui, Carlota O. [2] ; Pereira, Jorge F. B. [1]
Total Authors: 8
Affiliation:
[1] Sao Paulo State Univ UNESP, Sch Pharmaceut Sci, Dept Bioproc & Biotechnol, BR-14800903 Araraquara, SP - Brazil
[2] Univ Sao Paulo, Fac Ciencias Farmaceut, Dept Tecnol Bioquim Farmaceut, Sao Paulo, SP - Brazil
[3] Univ Aveiro, CICECO Aveiro Inst Mat, Dept Chem, Aveiro - Portugal
Total Affiliations: 3
Document type: Journal article
Source: JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY; v. 95, n. 4 DEC 2019.
Web of Science Citations: 0
Abstract

BACKGROUND l-Asparaginase (ASNase) is an important biopharmaceutical for the treatment of acute lymphoblastic leukemia (ALL); however, with some restrictions due to its high manufacturing costs. Aqueous biphasic systems (ABS) have been suggested as more economical platforms for the separation/purification of proteins, but a full understanding of the mechanisms behind the ASNase partition is still a major challenge. Polymer/salt-based ABS with different driving-forces (salting-out and hydrophilicity/hydrophobicity effects) were herein applied to control the partition of commercial ASNase. RESULTS The main results showed the ASNase partition to the salt- or polymer-rich phase depending on the ABS studied, with extraction efficiencies higher than 95%. For systems composed of inorganic salts, the ASNase partition was controlled by the polyethylene glycol (PEG) molecular weight used. Cholinium-salts-based ABS were able to promote a preferential ASNase partition to the polymer-rich phase using PEG-600 and to the salt-rich phase using a more hydrophobic polypropylene glycol (PPG)-400 polymer. It was possible to select the ABS composed of PEG-2000 + potassium phosphate buffer as the most efficient to separate the ASNase from the main contaminant proteins (purification factor = 2.4 +/- 0.2), while it was able to maintain the enzyme activity for posterior application as part of a therapeutic. CONCLUSION Polymer/salt ABS can be used to control the partition of ASNase and adjust its purification yields, demonstrating the ABS potential as more economic platform for the selective recovery of therapeutic enzymes from complex broths. (c) 2019 Society of Chemical Industry (AU)

FAPESP's process: 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)
Grantee:Jorge Fernando Brandão Pereira
Support type: Research Grants - Young Investigators Grants
FAPESP's process: 15/07749-2 - Protein engineering and comparison of microbial expression systems of the biopharmaceutical L-asparaginase
Grantee:Gisele Monteiro
Support type: Regular Research Grants
FAPESP's process: 13/08617-7 - Production of extracellular L-asparaginase: from bioprospecting to the engineering of an antileukemic biopharmaceutical
Grantee:Adalberto Pessoa Junior
Support type: Research Projects - Thematic Grants
FAPESP's process: 18/25994-2 - Development of novel platforms for pegylation of proteins with therapeutic potential using microfluidics
Grantee:João Henrique Picado Madalena Santos
Support type: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 18/15104-0 - Preclinical assays of glycoprotein asparaginase proteoforms or resistant to serum proteases
Grantee:Gisele Monteiro
Support type: Regular Research Grants
FAPESP's process: 14/19793-3 - Optimization and scale-up of novel Ionic-Liquid-based purification processes for recombinant green fluorescent protein produced by Escherichia coli "GFPurIL"
Grantee:Sandro Roberto Valentini
Support type: Regular Research Grants