Abstract
AimsThe aim of the current study was to develop a method to detect peptide-linked nanoparticles in blood plasma.Materials & MethodsA convenient enzyme linked immunosorbent assay (ELISA) was developed for the detection of peptides functionalized with biotin and fluorescein groups. As a proof of principle, polymerized pentafluorophenyl methacrylate nanoparticles linked to biotin-carboxyfluorescein labeled peptides were intravenously injected in Wistar rats. Serial blood plasma samples were analyzed by ELISA and by liquid chromatography mass spectrometry (LC/MS) technology.ResultsThe ELISA based method for the detection of FITC labeled peptides had a detection limit of 1 ng/mL. We were able to accurately measure peptides bound to pentafluorophenyl methacrylate nanoparticles in blood plasma of rats, and similar results were obtained by LC/MS.ConclusionsWe detected FITC-labeled peptides on pentafluorophenyl methacrylate nanoparticles after injection in vivo. This method can be extended to detect nanoparticles with different chemical compositions.
Highlights
The work presented here was developed by the FP6 EU biopharmaceutics platform, which aimed at the development of innovative multidisciplinary approaches for the design, synthesis and evaluation of molecular, nano- and micro-scale functionalities for targeted delivery of therapeutic peptides and proteins.The use of therapeutic peptides in neurodegenerative diseases is an active area of investigation
We were able to accurately measure peptides bound to pentafluorophenyl methacrylate nanoparticles in blood plasma of rats, and similar results were obtained by LC/MS
We used an enzyme linked immunosorbent assay (ELISA)-based method to detect the reporter peptide bound to the nanoparticles (Fig 1C), allowing quantification of these nanoparticles in blood plasma after injection in vivo
Summary
The work presented here was developed by the FP6 EU biopharmaceutics platform, which aimed at the development of innovative multidisciplinary approaches for the design, synthesis and evaluation of molecular, nano- and micro-scale functionalities for targeted delivery of therapeutic peptides and proteins.The use of therapeutic peptides in neurodegenerative diseases is an active area of investigation. Significant progress has been made towards organ-specific delivery of nanoparticles, a drawback is that they often do not reach their intended target tissue in the desired quantities due to filtering by the kidney, liver and spleen [7] This can be improved by decorating the nanoparticles with functionalized targeting peptides that bind to receptors on the target tissue (Fig 1) [13]. It is useful to have reliable methods based on commonly used laboratory techniques, to be able to measure nanoparticles in biological fluids and tissues In this manuscript we attached labeled reporter peptides to nanoparticles [6]. We used an ELISA-based method to detect the reporter peptide bound to the nanoparticles (Fig 1C), allowing quantification of these nanoparticles in blood plasma after injection in vivo. LC/MS analysis of the same samples was performed to measure the plasma concentration over time of acrylamide based nanoparticles loaded with reporter peptides in rats
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