Challenges in the analytical characterization of PEGylated asparaginase

Brandenburg, Gunda; Poppenborg, Sabine; Radcke, Christoph; Wittmann, Julia; Krähmer, Ralf; Pasut, Gianfranco; Leenders, Frank

doi:10.1016/b978-0-444-64081-9.00010-3

Cited by 3 publications

(4 citation statements)

References 28 publications

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“…On the other hand, a noticeable fraction of the conjugated subunits was of polyPEGylated derivatives (2-PEG, 11.3%; 3-PEG, 12.7%; and 4-PEG, 7.8%). ASNase has been PEGylated with different numbers of 5 kDa PEG chains: from 29 35 to 36−40 8 per tetrameric enzyme. In this work, a 12 kDa PEG was used.…”

Section: Resultsmentioning

confidence: 99%

“…In these cases, the protection provided by the PEG binding is lost. Moreover, the succinate spacer that remains in the protein after the hydrolysis can act as a hapten and may lead to an increased immunogenicity of the remaining protein. , Despite this problem, several biosimilars of pegaspargase have been developed and approved by regulatory agencies. ,, Currently, there are at least three L-ASNases available in the United States market: Oncaspar, Erwinaze, and Asparlas . Asparlas received its approval by the FDA in 2018, showing that the use of ASNase as a drug continues to be an active field of research and development…”

Section: Introductionmentioning

confidence: 99%

“…7 Pegaspargase is a conjugate of the ASNase of E. coli with 36− 40 PEG chains (5 kDa) with a succinic acid-based spacer. 8 The ester bond in the spacer between PEG and protein is relatively unstable and can be hydrolyzed under slightly alkaline conditions and, in vivo, by enzymatic cleavage. In these cases, the protection provided by the PEG binding is lost.…”

Section: Introductionmentioning

confidence: 99%

“…9,10 Despite this problem, several biosimilars of pegaspargase have been developed and approved by regulatory agencies. 8,11,12 Currently, there are at least three L-ASNases available in the United States market: Oncaspar, Erwinaze, and Asparlas. 13 Asparlas received its approval by the FDA in 2018, showing that the use of ASNase as a drug continues to be an active field of research and development.…”

Section: Introductionmentioning

confidence: 99%

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Integrated Lab-Scale Process Combining Purification and PEGylation of l-Asparaginase from Zymomonas mobilis

Ramón

Gonçalves

Alvarenga

et al. 2021

Ind. Eng. Chem. Res.

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During the last quarter of a century, PEGylation has been used to enhance the clinical effectiveness of various therapeutic proteins. However, the conjugation process significantly increases the costs of the process. This parameter is critical, especially for biosimilar products that do not have patent protection. The integration of protein purification to its conjugation could potentially reduce total costs. Following this hypothesis, in this work, the purification of L-asparaginase from Zymomonas mobilis, produced recombinantly in Escherichia coli, linked to its conjugation with a PEG of 12 kDa, was studied. The effect of operating parameters on cell rupture by high-pressure homogenization was analyzed, and the optimal values for the conditions in this study were determined. The chromatographic capture and final purification steps were also studied. It was possible to elute the protein from ion-exchange chromatography, used as final purification, in the optimal buffer for the PEGylation step. This result allowed us to eliminate the buffer change step that is usually carried out at the beginning of the PEGylation process. The conjugate obtained by this continuous process showed higher thermal stability and slower protein degradation compared to the native protein. This continuous process not only has great potential to reduce costs but could also be useful for studying the effect of different parameters on the overall performance of the process. This knowledge is especially important to establish a quality by design approach at the industrial scale.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Integrated Lab-Scale Process Combining Purification and PEGylation of l-Asparaginase from Zymomonas mobilis

Ramón

Gonçalves

Alvarenga

et al. 2021

Ind. Eng. Chem. Res.

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Modified Asparaginase for Treatment of Cancer Historical Appraisal and Future Perspectives

Icart

Araújo

Almeida

et al. 2023

CAPS

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Asparaginase (ASNase) is widely used as an important component of first-line treatment for acute lymphoblastic leukemia (ALL). Although it is associated with a high rate of complete remission (~93%), challenges remain due to several side effects ranging from immune reactions to severe toxicity, largely associated with its higher immunogenicity and glutamine coactivity. Innovative products have therefore been devised to minimise these adverse reactions while increasing the enzymes’ pharmacokinetic properties, stability, and efficacy. This review focuses on commercially available formulations and others that remain in development, discussing the most recent strategies for preparing alternative formulations of the enzyme to be less immunogenic and have low glutaminase coactivity by using site-specific mutagenesis, PEGylation, and encapsulation techniques.

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Cancer Nanomedicine and Immune System—Interactions and Challenges

Ahmad

Idris

Hanaffi

et al. 2021

Front. Nanotechnol.

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Nanoparticles have tremendous therapeutic potential in the treatment of cancer as they increase drug delivery, attenuate drug toxicity, and protect drugs from rapid clearance. Since Doxil®, the first FDA-approved nanomedicine, several other cancer nanomedicines have been approved and have successfully increased the efficacy over their free drug counterparts. Although their mechanisms of action are well established, their effects towards our immune system, particularly in the tumor microenvironment (TME), still warrant further investigation. Herein, we review the interactions between an approved cancer nanomedicine with TME immunology. We also discuss the challenges that need to be addressed for the full clinical potential of ongoing cancer nanomedicines despite the encouraging preclinical data.

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Challenges in the analytical characterization of PEGylated asparaginase

Cited by 3 publications

References 28 publications

Integrated Lab-Scale Process Combining Purification and PEGylation of l-Asparaginase from Zymomonas mobilis

Integrated Lab-Scale Process Combining Purification and PEGylation of l-Asparaginase from Zymomonas mobilis

Modified Asparaginase for Treatment of Cancer Historical Appraisal and Future Perspectives

Cancer Nanomedicine and Immune System—Interactions and Challenges

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