Detection of Peptide-based nanoparticles in blood plasma by ELISA

PLoS One. 2015 May 21;10(5):e0126136. doi: 10.1371/journal.pone.0126136. eCollection 2015.

Abstract

Aims: The aim of the current study was to develop a method to detect peptide-linked nanoparticles in blood plasma.

Materials & methods: A 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.

Results: The 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.

Conclusions: We detected FITC-labeled peptides on pentafluorophenyl methacrylate nanoparticles after injection in vivo. This method can be extended to detect nanoparticles with different chemical compositions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Liquid
  • Enzyme-Linked Immunosorbent Assay* / methods
  • Male
  • Nanoparticles*
  • Peptides* / blood
  • Plasma*
  • Rats
  • Tandem Mass Spectrometry

Substances

  • Peptides

Grants and funding

This work was supported by a grant from the EC-FP6 (to the European NanoBioPharmaceutics research initiative). AplaGen GmbH provided support in the form of salaries for authors Andy J.G. Pötgens and Hans-Georg Frank, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.