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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Novel site-specific PEGylated L-asparaginase

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Autor(es):
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Meneguetti, Giovanna Pastore [1] ; Picado Madalena Santos, Joao Henrique [1, 2] ; Torres Obreque, Karin Mariana [1] ; Vaz Barbosa, Christiano Marcello [3] ; Monteiro, Gisele [1] ; Poliselli Farsky, Sandra Helena [3] ; de Oliveira, Adriano Marim [4] ; Angeli, Claudia Blanes [5] ; Palmisano, Giuseppe [5] ; Marques Ventura, Sonia Patricia [2] ; Pessoa-Junior, Adalberto [1] ; Rangel-Yagui, Carlota de Oliveira [1]
Número total de Autores: 12
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Dept Biochem & Pharmaceut Technol, Sao Paulo - Brazil
[2] Univ Aveiro, Dept Chem, CICECO Aveiro Inst Mat, Aveiro - Portugal
[3] Univ Sao Paulo, Dept Clin & Toxicol Anal, Sao Paulo - Brazil
[4] Technol Res Inst, Dept Bionanomanufacture, Sao Paulo - Brazil
[5] Univ Sao Paulo, Inst Biomed Sci, Dept Parasitol, Sao Paulo - Brazil
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: PLoS One; v. 14, n. 2 FEB 12 2019.
Citações Web of Science: 1
Resumo

L-asparaginase (ASNase) from Escherichia coli is currently used in some countries in its PEGylated form (ONCASPAR, pegaspargase) to treat acute lymphoblastic leukemia (ALL). PEGylation refers to the covalent attachment of poly(ethylene) glycol to the protein drug and it not only reduces the immune system activation but also decreases degradation by plasmatic proteases. However, pegaspargase is randomly PEGylated and, consequently, with a high degree of polydispersity in its final formulation. In this work we developed a site-specific N-terminus PEGylation protocol for ASNase. The monoPEG-ASNase was purified by anionic followed by size exclusion chromatography to a final purity of 99%. The highest yield of monoPEG-ASNase of 42% was obtained by the protein reaction with methoxy polyethylene glycol-carboxymethyl N-hydroxysuccinimidyl ester (10kDa) in 100 mM PBS at pH 7.5 and PEG: ASNase ratio of 25:1. The monoPEG-ASNase was found to maintain enzymatic stability for more days than ASNase, also was resistant to the plasma proteases like asparaginyl endopeptidase and cathepsin B. Additionally, monoPEG-ASNase was found to be potent against leukemic cell lines (MOLT-4 and REH) in vitro like polyPEG-ASNase. monoPEG-ASNase demonstrates its potential as a novel option for ALL treatment, being an inventive novelty that maintains the benefits of the current enzyme and solves challenges. (AU)

Processo FAPESP: 16/22065-5 - Pegilação N-terminal de proteínas e purificação por sistemas aquosos bifásicos
Beneficiário:Carlota de Oliveira Rangel Yagui
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 13/08617-7 - Produção de L-asparaginase extracelular: da bioprospecção à engenharia de um biofármaco antileucêmico
Beneficiário:Adalberto Pessoa Junior
Linha de fomento: Auxílio à Pesquisa - Temático