title: Quantification of Cell-Penetrating Peptide Associated with Polymeric Nanoparticles Using Isobaric-Tagging and MALDI-TOF MS/MS.
abstract: High sensitivity quantification of the putative cell-penetrating peptide di-arginine-histidine (RRH) associated with poly (ethyl-cyanoacrylate) (PECA) nanoparticles was achieved without analyte separation, using a novel application of isobaric-tagging and high matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometry. Isobaric-tagging reaction equilibrium was reached after 5 min, with 90% or greater RRH peptide successfully isobaric-tagged after 60 min. The accuracy was greater than 90%, which indicates good reliability of using isobaric-tagged RRH as an internal standard for RRH quantification. The sample intra- and inter-spot coefficients of variations were less than 11%, which indicate good repeatability. The majority of RRH peptides in the nanoparticle formulation were physically associated with the nanoparticles (46.6%), whereas only a small fraction remained unassociated (13.7%). The unrecovered RRH peptide (~40%) was assumed to be covalently associated with PECA nanoparticles. Graphical Abstract ᅟ.
authors.affiliation: School of Pharmacy, University of Otago, 18 Frederick Street, Dunedin, 9016, New Zealand.
authors.forename: Jasper Z S
authors.initials: JZ
authors.lastname: Chiu
authors.affiliation: School of Pharmacy, University of Otago, 18 Frederick Street, Dunedin, 9016, New Zealand.
authors.forename: Ian G
authors.initials: IG
authors.lastname: Tucker
authors.affiliation: School of Pharmacy, University of Otago, 18 Frederick Street, Dunedin, 9016, New Zealand. arlene.mcdowell@otago.ac.nz.
authors.forename: Arlene
authors.initials: A
authors.lastname: McDowell
mesh_terms: D057846:Cell-Penetrating Peptides / Q000032:analysis*; D053758:Nanoparticles; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D057846:Cell-Penetrating Peptides
keywords: Isobaric-tagging; MALDI-TOF; Nanoparticles; Oligoarginine; Peptide quantification; Quantification