Critical overview of the main features and techniques used for the evaluation of the clinical applicability of L-asparaginase as a biopharmaceutical to treat blood cancer

Costa-Silva, Tales A.; Costa, Iris Munhoz; Biasoto, Henrique Pellin; Lima, Guilherme Meira; Silva, C.; Pessoa, Adalberto; Monteiro, Gisele

doi:10.1016/j.blre.2020.100651

Cited by 43 publications

(36 citation statements)

References 154 publications

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“…It was only in 2006 that it was approved as part of the first-line therapy for any ALL patient [57]. In November 2011, FDA approved Erwinase ® , indicating its use as a component of a multi-agent chemotherapeutic regimen for ALL patients treatment who have developed hypersensitivity to either Elspar ® or Oncaspar ® [58]. Finally, in January 2016, the European Commission granted a marketing authorisation valid throughout the European Union for Spectrila ® (from E. coli).…”

Section: Commercial Asnasementioning

confidence: 99%

Recent Strategies and Applications for l-Asparaginase Confinement

et al. 2020

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l-asparaginase (ASNase, EC 3.5.1.1) is an aminohydrolase enzyme with important uses in the therapeutic/pharmaceutical and food industries. Its main applications are as an anticancer drug, mostly for acute lymphoblastic leukaemia (ALL) treatment, and in acrylamide reduction when starch-rich foods are cooked at temperatures above 100 °C. Its use as a biosensor for asparagine in both industries has also been reported. However, there are certain challenges associated with ASNase applications. Depending on the ASNase source, the major challenges of its pharmaceutical application are the hypersensitivity reactions that it causes in ALL patients and its short half-life and fast plasma clearance in the blood system by native proteases. In addition, ASNase is generally unstable and it is a thermolabile enzyme, which also hinders its application in the food sector. These drawbacks have been overcome by the ASNase confinement in different (nano)materials through distinct techniques, such as physical adsorption, covalent attachment and entrapment. Overall, this review describes the most recent strategies reported for ASNase confinement in numerous (nano)materials, highlighting its improved properties, especially specificity, half-life enhancement and thermal and operational stability improvement, allowing its reuse, increased proteolysis resistance and immunogenicity elimination. The most recent applications of confined ASNase in nanomaterials are reviewed for the first time, simultaneously providing prospects in the described fields of application.

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Section: Commercial Asnasementioning

confidence: 99%

Recent Strategies and Applications for l-Asparaginase Confinement

et al. 2020

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“…We purified p3-ScA5 and p8-ScA50 from the excess of unreacted ScA using PEG-NaCl precipitation, 31 and confirmed the removal of the free ScA by SDS-PAGE ( Figure 1c). The commonly used colorimetric Nessler assay 38 demonstrated that both p3-and p8-constructs were enzymatically active (Figure 2a). Mixing the ScA sample with a phage that display the random SVEK peptide sequence or precipitation of p3-st and p8-st with no ScA presented, resulted in samples with no enzymatic activity (Figure 2a).…”

Section: Conjugation Of Erwinase To Spytag-displaying Phagesmentioning

confidence: 99%

DNA-Encoded Multivalent Display of Protein Tetramers on Phage: Synthesis and In Vivo Aplications

Lima

Atrazhev

Sarkar

et al. 2021

Preprint

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Phage display links phenotype of displayed polypeptides with DNA sequence in phage genome and offers a universal method for discovery of proteins with novel properties. Injection of phage-displayed libraries in living organisms further provides a unique and powerful approach to optimize biochemical, pharmacological and biological properties of the displayed peptides, antibodies and other proteins in vivo. However, over 60% of the proteome is comprised of multi-domain proteins, and display of large multi-subunit proteins on phages remains a challenge. Majority of protein display systems are based on monovalent phagemid constructs but methods for robust display of multiple copies of large proteins are scarce. Here, we describe a DNA-encoded display of a ~200 kDa tetrameric protein tetrameric L-asparaginase on M13 phage produced by ligation of SpyCatcher-Asparaginase fusion (ScA) to prospectively barcoded phage clones displaying SpyTag peptide. Starting from the SpyTag display on p3 minor coat protein or p8 major coat protein yielded constructs with five copies of ScA displayed on p3 (ScA5-phage) and 50 copies of ScA on p8 protein (ScA50-phage). ScA remained active after conjugation. It could be easily produced directly from lysates of bacteria that express ScA. Display constructs of different valency can be injected into mice and analyzed by deep-sequencing of the DNA barcodes associated phage clones. In these multiplexed studies, we observed a density-dependent clearance rate in vivo. A known clearance mechanism of L-asparaginase is endocytosis by phagocytic cells. Our observations, thus, link the increase in density of the displayed protein with the increased rate of the endocytosis by cells in vivo. In conclusion, we demonstrate that a multivalent display of L-asparaginase on phage could be used to study the circulation life of this protein in vivo and such approach opens the possibility to use DNA sequencing to investigate multiplexed libraries of other multi-subunit proteins in vivo.

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“…Currently, ASNase is obtained from prokaryotic microorganisms such as Escherichia coli and Erwinia chrysanthemi 4 . However, when administered in humans, ASNase can cause severe immunological reactions often resulting in the interruption of treatment 5,6 .…”

Section: Introductionmentioning

confidence: 99%

“…3 Currently, ASNase is obtained from prokaryotic microorganisms such as Escherichia coli and Erwinia chrysanthemi. 4 However, when administered in humans, ASNase can cause severe immunological reactions often resulting in the interruption of treatment. 5,6 The ASNase of E. coli is particularly hyperallergenic in humans because of its high molecular weight (the active tetramer is 140 kDa) and its amino acid sequence that contains immunogenic regions.…”

Section: Introductionmentioning

confidence: 99%

Recombinant l‐asparaginase production using Pichia pastoris (MUT^s strain): establishment of conditions for growth and induction phases

Pillaca-Pullo

Rodrigues

Sánchez‐Moguel

et al. 2020

J of Chemical Tech & Biotech

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BACKGROUND: L-asparaginase (ASNase), a biopharmaceutical enzyme used in the treatment of childhood lymphoid malignancies, is commercially produced from Escherichia coli and Erwinia chrysanthemi. However, it causes severe adverse effects due to allergenic prokaryotic epitopes on the protein surface. ASNase II from Saccharomyces cerevisiae can be a promising alternative source of this enzyme. In this study, conditions to produce ASNase from S. cerevisiae expressed in Pichia pastoris have been investigated in shake flasks and 3 L-bioreactor. We evaluated if medium composition, concentration of carbon source (i.e. glycerol), growth time, concentration of inducer (i.e. methanol), temperature and initial pH influenced both biomass and ASNase expression. RESULTS: Biomass of around 53 g L-1 and ASNase volumetric activity of 710 U L-1 were achieved using the buffered glycerol-complex medium (BMGY) containing 40 g L-1 glycerol, with induction after 141 h using 3.0% (v/v) methanol, at 20°C and initial pH 6.0. CONCLUSION: The experiments performed in shake flasks were scalable to a 3 L-bioreactor, suggesting that this bioprocess could be scaled-up for industrial production.

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Critical overview of the main features and techniques used for the evaluation of the clinical applicability of L-asparaginase as a biopharmaceutical to treat blood cancer

Cited by 43 publications

References 154 publications

Recent Strategies and Applications for l-Asparaginase Confinement

Recent Strategies and Applications for l-Asparaginase Confinement

DNA-Encoded Multivalent Display of Protein Tetramers on Phage: Synthesis and In Vivo Aplications

Recombinant l‐asparaginase production using Pichia pastoris (MUT^s strain): establishment of conditions for growth and induction phases

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Critical overview of the main features and techniques used for the evaluation of the clinical applicability of L-asparaginase as a biopharmaceutical to treat blood cancer

Cited by 43 publications

References 154 publications

Recent Strategies and Applications for l-Asparaginase Confinement

Recent Strategies and Applications for l-Asparaginase Confinement

DNA-Encoded Multivalent Display of Protein Tetramers on Phage: Synthesis and In Vivo Aplications

Recombinant l‐asparaginase production using Pichia pastoris (MUTs strain): establishment of conditions for growth and induction phases

Contact Info

Product

Resources

About

Recombinant l‐asparaginase production using Pichia pastoris (MUT^s strain): establishment of conditions for growth and induction phases