Development and analysis of beta glucan particles for high throughput peptide and protein loading Public
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The goal of this project was to develop high-throughput protein and peptide loading of beta glucan particles (GPs) for vaccine development in 96-well plates. After establishing ideal loading conditions to be similar to those of standard GP loading, peptide binding using N-succinimidyl S-acetylthioacetate (SATA) and a new binding method using 2,2'-dithiolethyl bis(4-azido-2,3,5,6-tetrafluorobenzoate) (DAT) showed that the DAT GPs bound peptide more efficiently than the SATA GPs. GP-mediated peptide delivery to OT-II T-cells showed that DAT-derivatized GPs stimulated a stronger immune response than linear PEI/SATA-derivatized GPs. Immunological testing for Ova protein-loaded GPs versus IgG- and lysozyme-loaded GPs showed that both Ova protein- and IgG-loaded GPs stimulated a T-cell response.
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