Engineering Cell-Free Protein Synthesis for High-Yield Production and Human Serum Activity Assessment of Asparaginase: Toward On-Demand Treatment of Acute Lymphoblastic Leukemia.
Antineoplastic Agents
/ therapeutic use
Asparaginase
/ biosynthesis
Bacteria
/ enzymology
Batch Cell Culture Techniques
/ methods
Cell Engineering
Escherichia coli
/ metabolism
Humans
Precursor Cell Lymphoblastic Leukemia-Lymphoma
/ drug therapy
Protein Biosynthesis
Protein Engineering
/ methods
Serum
/ enzymology
asparaginase assay
cell-free protein synthesis
crisantaspase
human serum
lymphoblastic leukemia
Journal
Biotechnology journal
ISSN: 1860-7314
Titre abrégé: Biotechnol J
Pays: Germany
ID NLM: 101265833
Informations de publication
Date de publication:
Apr 2020
Apr 2020
Historique:
received:
02
07
2019
revised:
13
11
2019
pubmed:
7
1
2020
medline:
27
10
2020
entrez:
7
1
2020
Statut:
ppublish
Résumé
Acute lymphocytic leukemia (ALL) is a common childhood cancer in the United States, with over 6000 new cases diagnosed each year. Administration of bacterial asparaginase (ASNase) has improved survival rates to nearly 80%, however these therapeutics have high incidence of immunological neutralization and serum activity must be monitored for most effective treatment regimens. Here, a 72% improvement in cell-free protein synthesis (CFPS) of FDA approved l-asparaginase (crisantaspase) is demonstrated by employing an aspartate-fed-batch reactor format. A CFPS-based ASNase activity assay as a tool for therapeutic regimentation and production quality control is also presented. This work suggests that shelf-stable and low-cost Escherichia coli-based CFPS reactions may be employed on-demand to 1) synthesize biologics on-site for patient administration, 2) verify biologic activity for dosage calculations, and 3) monitor therapeutic activity in human serum during the treatment regimen. The combination of both therapeutic production and activity assessment introduces a concept of synergistic utility for bacterial cell lysates in modern medical treatment. Indeed, recent work with CFPS biosensors supports a not-too-distant future when shelf-stable E. coli CFPS systems are used to diagnose, treat, and monitor treatment of diseases in the clinical setting.
Identifiants
pubmed: 31904183
doi: 10.1002/biot.201900294
doi:
Substances chimiques
Antineoplastic Agents
0
Asparaginase
EC 3.5.1.1
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e1900294Subventions
Organisme : Division of Chemical, Bioengineering, Environmental, and Transport Systems
ID : 1254148
Organisme : Brigham Young University
ID : Simmons Center for Cancer Research Fellowship
Informations de copyright
© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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