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Biotechnol Prog. 2012 Mar-Apr;28(2):567-72. doi: 10.1002/btpr.1516. Epub 2012 Feb 17.

Expression of a cGMP compatible Lucilia sericata insect serine proteinase debridement enzyme.

Author information

1
Immune Modulation Research Group, School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, United Kingdom. david.pritchard@nottingham.ac.uk

Abstract

Previously, we demonstrated the effectiveness of a research grade recombinant chymotrypsin, derived from the larvae of Lucilia sericata, in "debriding" slough/eschar from venous leg ulcers ex vivo. Furthermore, we were able to formulate this enzyme for successful delivery to in vitro wound healing assays, from a prototype hydrogel wound dressing, and showed that enzyme delivered in this way could degrade wound tissue ex vivo. Recently, to progress biotechnological development of the enzyme as a potential therapeutic product, we explored expression using current good manufacturing practice (cGMP) guidelines, and now report that a recombinant chymotrypsin I zymogen from L. sericata can be expressed in the cGMP acceptable strain of Escherichia coli (BLR-DE3). In addition, the conditions required for purification, refolding and activation of the chymotrypsinogen have been determined. The activated enzyme was stable, and effective in digesting wound slough/eschar tissue. To summarise, we have successfully initiated the production and characterisation of a novel cGMP compatible product for use in future clinical trials.

PMID:
22323388
DOI:
10.1002/btpr.1516
[Indexed for MEDLINE]

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