Prevention of Aggregation and Autocatalysis for Sustaining Biological Activity of Recombinant BoNT/A-LC Upon Long-Term Storage

Author(s): Padma Singh, Manglesh Kumar Singh, Vinita Chauhan, Pallavi Gupta, Ram Kumar Dhaked.

Journal Name: Protein & Peptide Letters

Volume 18 , Issue 12 , 2011

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Abstract:

Protein aggregation during expression, purification, storage, or transfer into requisite assay buffers hampers the use of proteins for in vitro studies. The formation of these aggregates represents a major obstacle in the study of biological activity and also restricts the spectrum of protein products being available for the biomedical applications. The catalytic light chain of botulinum neurotoxin type A undergoes autocatalysis and aggregation after purification upon long-term storage and freeze-thawing. In present study the conditions for the high level expression and purification of biologically active light chain protein of botulinum neurotoxin were optimized from a synthetic gene. Several co-solvents were screened in order to prevent autocatalysis and aggregation of rBoNT/A-LC. The effect of the co-solvents is studied on endopeptidase activity during long term storage of the recombinant protein. The purified rBoNT/A-LC was also evaluated for its immunogenicity.

Keywords: Botulinum neurotoxin, SNAP-25, endopeptidase, autocatalysis, aggregation, glycerol, LC, HC, BoNT, rBoNT/A-LC, VAMP, MALDI-TOF, HRP, HUPOBotulinum neurotoxin, SNAP-25, endopeptidase, autocatalysis, aggregation, glycerol, LC, HC, BoNT, rBoNT/A-LC, VAMP, MALDI-TOF, HRP, HUPO

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Article Details

VOLUME: 18
ISSUE: 12
Year: 2011
Page: [1177 - 1187]
Pages: 11
DOI: 10.2174/092986611797642689
Price: $65

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