Monitoring the Early Stage Self-Assembly of Enzyme-Assisted Peptide Hydrogels
Richard J. Williams A B , James Gardiner A C , Anders B. Sorensen A , Silvia Marchesan A , Roger J. Mulder A , Keith M. McLean A and Patrick G. Hartley AA CSIRO Materials Science and Engineering, Clayton, Vic. 3168, Australia.
B School of Life and Enviromental Sciences, Deakin University, Waurn Ponds, Vic. 3026, Australia.
C Corresponding author. Email: james.gardiner@csiro.au
Australian Journal of Chemistry 66(5) 572-578 https://doi.org/10.1071/CH12557
Submitted: 19 December 2012 Accepted: 24 January 2013 Published: 21 February 2013
Abstract
The early stages of the self-assembly of peptide hydrogels largely determine their final material properties. Here we discuss experimental methodologies for monitoring the self-assembly kinetics which underpin peptide hydrogel formation. The early stage assembly of an enzyme-catalysed Fmoc-trileucine based self-assembled hydrogel was examined using spectroscopic techniques (circular dichroism, CD, and solution NMR) as well as chromatographic (HPLC) and mechanical (rheology) techniques. Optimal conditions for enzyme-assisted hydrogel formation were identified and the kinetics examined. A lag time associated with the formation and accumulation of the self-assembling peptide monomer was observed and a minimum hydrogelator concentration required for gelation was identified. Subsequent formation of well defined nano- and microscale structures lead to self-supporting hydrogels at a range of substrate and enzyme concentrations. 1H NMR monitoring of the early self-assembly process revealed trends that were well in agreement with those identified using traditional methods (i.e. HPLC, CD, rheology) demonstrating 1H NMR spectroscopy can be used to non-invasively monitor the self-assembly of peptide hydrogels without damaging or perturbing the system.
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