Novel site-specific PEGylated L-asparaginase
authors Meneguetti, GP; Santos, JHPM; Obreque, KMT; Barbosa, CMV; Monteiro, G; Farsky, SHP; de Oliveira, AM; Angeli, CB; Palmisano, G; Ventura, SPM; Pessoa, A; Rangel-Yagui, CD
nationality International
journal PLOS ONE
keywords ESCHERICHIA-COLI ASPARAGINASE; SUBSTRATE-SPECIFICITY; PURIFICATION; PROTEINS; LEUKEMIA; PEGASPARGASE; SYNTHETASE; EXPRESSION; RESISTANCE; CHILDREN
abstract 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.
publisher PUBLIC LIBRARY SCIENCE
issn 1932-6203
year published 2019
volume 14
issue 2
digital object identifier (doi) 10.1371/journal.pone.0211951
web of science category Multidisciplinary Sciences
subject category Science & Technology - Other Topics
unique article identifier WOS:000458395600031
  ciceco authors
  impact metrics
journal analysis (jcr 2017):
journal impact factor 2.766
5 year journal impact factor 3.352
category normalized journal impact factor percentile 77.344
dimensions (citation analysis):
altmetrics (social interaction):



 


Sponsors

1suponsers_list_ciceco.jpg