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

Santosh Gurram, Gopal Srivastava, Vishnupriya Badve, Vinod Nandre, Shridevi Gundu, Pooja Doshi

Applied biochemistry and biotechnology 2022 Feb

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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.

Citation

Santosh Gurram, Gopal Srivastava, Vishnupriya Badve, Vinod Nandre, Shridevi Gundu, Pooja Doshi. Pyridine Borane as Alternative Reducing Agent to Sodium Cyanoborohydride for the PEGylation of L-asparaginase. Applied biochemistry and biotechnology. 2022 Feb;194(2):827-847