Detection of Peptide-based nanoparticles in blood plasma by ELISA.
Gerard H Bode, Karin E Pickl, Maria Sanchez-Purrà, Berta Albaiges, Salvador Borrós, Andy J G Pötgens, Christoph Schmitz, Frank M Sinner, Mario Losen, Harry W M Steinbusch, Hans-Georg Frank, Pilar Martinez-Martinez
The aim of the current study was to develop a method to detect peptide-linked nanoparticles in blood plasma.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.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.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.
