Improving peptide quantification in chitosan nanoparticles

The objective of the present study was to evaluate different methodologies for peptide quantification in the supernatant of chitosan nanoparticles by removing the unliked polymer in the suspension. The ionic gelation method was used to prepare the chitosan nanoparticles encapsulating a 5.3 kDa peptide. Three different methodologies for the processment of the solutions were compared before subjecting the samples to the Bradford protocol or Qubit® kit for protein detection. For the quantification, it was necessary to create a standard peptide curve using different peptide concentrations. The suitable standard curve would be one in which the peptide was diluted in the empty chitosan supernatant (obtained after nanoparticles centrifugation) or in the filtrate of the empty chitosan supernatant. Our results indicated the safest methodology tested for peptide quantification in the supernatant chitosan nanoparticles was filtering the solution before subjecting the sample to the Bradford assay.