Pyridine Borane as Alternative Reducing Agent to Sodium Cyanoborohydride for the PEGylation of L-asparaginase.

Autor: Gurram S; Division of Biochemistry, Department of Chemistry, Savitribai Phule Pune University, Formerly University of Pune), Ganeshkhind, Pune, 411007, Maharashtra, India., Srivastava G; Division of Biochemistry, Department of Chemistry, Savitribai Phule Pune University, Formerly University of Pune), Ganeshkhind, Pune, 411007, Maharashtra, India., Badve V; Division of Biochemistry, Department of Chemistry, Savitribai Phule Pune University, Formerly University of Pune), Ganeshkhind, Pune, 411007, Maharashtra, India., Nandre V; Division of Biochemistry, Department of Chemistry, Savitribai Phule Pune University, Formerly University of Pune), Ganeshkhind, Pune, 411007, Maharashtra, India., Gundu S; Division of Biochemistry, Department of Chemistry, Savitribai Phule Pune University, Formerly University of Pune), Ganeshkhind, Pune, 411007, Maharashtra, India., Doshi P; Division of Biochemistry, Department of Chemistry, Savitribai Phule Pune University, Formerly University of Pune), Ganeshkhind, Pune, 411007, Maharashtra, India. pooja.doshi@unipune.ac.in.
Jazyk: angličtina
Zdroj: Applied biochemistry and biotechnology [Appl Biochem Biotechnol] 2022 Feb; Vol. 194 (2), pp. 827-847. Date of Electronic Publication: 2021 Sep 22.
DOI: 10.1007/s12010-021-03657-y
Abstrakt: PEGylation is a reductive alkylation of a protein N-terminal/α-amine of protein with mPEG chain by reducing agent. To obtain quantitative and site-specific PEGylation, sodium cyanoborohydride is commonly used as a reducing agent. The reduction process of sodium cyanoborohydride produces highly poisonous hydrogen cyanide, which may render the final product toxic. Herein, we have studied various reducing agents such as dimethylamine borane, triethylamine borane, trimethylamine borane, pyridine borane, morpholine borane, 2-picoline borane, and 5-ethyl-2-methyl-pyridine borane were tested as alternatives to sodium cyanoborohydride for the PEGylation of L-asparaginase. The characterization of reacted pegaspargase was carried out by SDS-PAGE, Western blotting, SEC-HPLC, RP-HPLC, SEC-MALS, CD, enzyme activity, and cell proliferation assays using with lymphoblast cells and MTS/PMS as substrate. Pyridine borane was determined to be the best acceptable reducing agent for PEGylation in terms of purity and activity. As a result, instead of sodium cyanoborohydride, pyridine borane can be employed.
(© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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