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Journal Articles Analytical and Bioanalytical Chemistry Year : 2015

Real-time monitoring of peptide grafting onto chitosan films using capillary electrophoresis

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Danielle L. Taylor
  • Function : Author
Joel J. Thevarajah
  • Function : Author
Diksha K. Narayan
  • Function : Author
Patricia Murphy
  • Function : Author
Melissa M. Mangala
  • Function : Author
Seakcheng Lim
  • Function : Author
Richard Wuhrer
  • Function : Author
Catherine Lefay
Michael D. O'Connor
  • Function : Author
Patrice Castignolles

Abstract

Chitosan, being antimicrobial and biocompatible, is attractive as a cell growth substrate. To improve cell attachment, arginine-glycine-aspartic acid-serine (RGDS) peptides were covalently grafted to chitosan films, through the widely used coupling agents 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC-HCl) and N-hydroxysuccinimide (NHS), via the carboxylic acid function of the RGDS molecule. The grafting reaction was monitored, for the first time, in real time using free-solution capillary electrophoresis (CE). This enabled fast separation and determination of the peptide and all other reactants in one separation with no sample preparation. Covalent RGDS peptide grafting onto the chitosan film surface was demonstrated using solid-state NMR of swollen films. CE indicated that oligomers of RGDS, not simply RGDS, were grafted on the film, with a likely hyperbranched structure. To assess the functional properties of the grafted films, cell growth was compared on control and peptide-grafted chitosan films. Light microscopy and polymerase chain reaction (PCR) analysis demonstrated greatly improved cell attachment to RGDS-grafted chitosan films.
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Dates and versions

hal-01460653 , version 1 (07-02-2017)

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Danielle L. Taylor, Joel J. Thevarajah, Diksha K. Narayan, Patricia Murphy, Melissa M. Mangala, et al.. Real-time monitoring of peptide grafting onto chitosan films using capillary electrophoresis. Analytical and Bioanalytical Chemistry, 2015, 407 (9), pp.2543--2555. ⟨10.1007/s00216-015-8483-y⟩. ⟨hal-01460653⟩
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