Purification of Recombinant <i>L</i>‐Asparaginase II Using Solvent‐Freeze‐Out Technology

Yu, Xiaohua; Wu, Yining; Huang, Fang; Ulrich, Joachim; Wang, Jingkang

doi:10.1002/ceat.201700569

Cited by 6 publications

(2 citation statements)

References 24 publications

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“…Yu et al (2018) studied the purification of recombinant ASNase from E. coli using crystallization by solvent freezing technology (SFO) with methanol, 2-methyl-2,4-pentanediol (MPD), polyethylene glycol (PEG) 6,000 and ethanol as precipitating agents. Initially, the biomass was disintegrated via high pressure homogenization (1,000 bar).…”

Section: L-asparaginase Manufacturing: Production Process and Purificmentioning

confidence: 99%

Development of L-Asparaginase Biobetters: Current Research Status and Review of the Desirable Quality Profiles

Brumano

Silva

Costa-Silva

et al. 2019

Front. Bioeng. Biotechnol.

142

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L-Asparaginase (ASNase) is a vital component of the first line treatment of acute lymphoblastic leukemia (ALL), an aggressive type of blood cancer expected to afflict over 53,000 people worldwide by 2020. More recently, ASNase has also been shown to have potential for preventing metastasis from solid tumors. The ASNase treatment is, however, characterized by a plethora of potential side effects, ranging from immune reactions to severe toxicity. Consequently, in accordance with Quality-by-Design (QbD) principles, ingenious new products tailored to minimize adverse reactions while increasing patient survival have been devised. In the following pages, the reader is invited for a brief discussion on the most recent developments in this field. Firstly, the review presents an outline of the recent improvements on the manufacturing and formulation processes, which can severely influence important aspects of the product quality profile, such as contamination, aggregation and enzymatic activity. Following, the most recent advances in protein engineering applied to the development of biobetter ASNases (i.e., with reduced glutaminase activity, proteolysis resistant and less immunogenic) using techniques such as site-directed mutagenesis, molecular dynamics, PEGylation, PASylation and bioconjugation are discussed. Afterwards, the attention is shifted toward nanomedicine including technologies such as encapsulation and immobilization, which aim at improving ASNase pharmacokinetics. Besides discussing the results of the most innovative and representative academic research, the review provides an overview of the products already available on the market or in the latest stages of development. With this, the review is intended to provide a solid background for the current product development and underpin the discussions on the target quality profile of future ASNase-based pharmaceuticals.

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Section: L-asparaginase Manufacturing: Production Process and Purificmentioning

confidence: 99%

Development of L-Asparaginase Biobetters: Current Research Status and Review of the Desirable Quality Profiles

Brumano

Silva

Costa-Silva

et al. 2019

Front. Bioeng. Biotechnol.

142

View full text Add to dashboard Cite

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“…Since neither compound was incorporated as soaking/co-crystallization, the L-Asp was part of the product formulation. We found both mannitol and PEG in Leuginase, which could be present due to purification by crystallization [47] or to increase the stability of the formulation. The product leaflet, however, does not disclose any adjuvants in the formulation.…”

Section: Discussionmentioning

confidence: 99%

Biophysical characterization of two commercially available preparations of the drug containingEscherichia coliL-Asparaginase 2

Araújo

Scapin

Andrade

et al. 2020

Preprint

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The hydrolysis of asparagine and glutamine by L-asparaginase has been used to treat acute lymphoblastic leukemia for over four decades. Each L-asparaginase monomer has a long loop that closes over the active site upon substrate binding, acting as a lid. Here we present a comparative study two commercially available preparations of the drug containing Escherichia coli L-Asparaginase 2, performed by a comprehensive array of biophysical and biochemical approaches. We report the oligomeric landscape and conformational and dynamic plasticity of E. coli type 2 L-asparaginase (EcA2) present in two different formulations, and its relationship with L-aspartic acid, which is present in Aginasa, but not in Leuginase. EcA2 shows a composition of monomers and oligomers up to tetramers, which is mostly not altered in the presence of L-Asp. The N-terminal loop of Leuginase, which is part of the active site is flexibly disordered, but gets ordered as in Aginasa in the presence os L-Asp, while L-Glu only does so to a limited extent. Ion-mobility spectrometry–mass spectrometry reveals two conformers for the monomeric EcA2, one of which can selectively bind to L-Asp and L-Glu. Aginasa has higher resistance to in vitro proteolysis than Leuginase, and this is directly related to the presence of L-Asp.

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Applications of high-pressure homogenization on microbial enzymes

Aguilar¹

2023

Effect of High-Pressure Technologies on Enzymes

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Purification of Recombinant L‐Asparaginase II Using Solvent‐Freeze‐Out Technology

Cited by 6 publications

References 24 publications

Development of L-Asparaginase Biobetters: Current Research Status and Review of the Desirable Quality Profiles

Development of L-Asparaginase Biobetters: Current Research Status and Review of the Desirable Quality Profiles

Biophysical characterization of two commercially available preparations of the drug containingEscherichia coliL-Asparaginase 2

Applications of high-pressure homogenization on microbial enzymes

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