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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY; v. 95, n. 4, p. 1016-1027, APR 2020.
• Citações Web of Science: 3

Resumo

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)

• Processo FAPESP: 13/08617-7 - Produção de L-asparaginase extracelular: da bioprospecção à engenharia de um biofármaco antileucêmico
• Beneficiário: Adalberto Pessoa Junior
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/15104-0 - Ensaios pré-clínicos de proteoformas de asparaginase glicoproteicas ou resistentes a proteases séricas.
• Beneficiário: Gisele Monteiro
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/16424-7 - Otimização e aumento de escala do processo de extração líquido-líquido com líquidos iônicos (LIs) como ferramenta de separação sustentável do biofármaco antileucêmico L-asparaginase (ASPase)
• Beneficiário: Jorge Pereira
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 18/25994-2 - Desenvolvimento de novas plataformas de PEGuilação de proteínas com potencial terapêutico com recurso à microfluídica
• Beneficiário: João Henrique Picado Madalena Santos
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/19793-3 - Otimização e aumento de escala de processos com líquidos iônicos como nova ferramenta de purificação da proteína verde fluorescente recombinante produzida por Escherichia coli "GFPurIL"
• Beneficiário: Sandro Roberto Valentini
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/07749-2 - Engenharia de proteínas e comparação de sistemas microbianos de expressão do biofármaco L-asparaginase
• Beneficiário: Gisele Monteiro
• Modalidade de apoio: Auxílio à Pesquisa - Regular