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Detection of Peptide-based nanoparticles in blood plasma by ELISA.

Bode GH, Pickl KE, Sanchez-Purrà M, Albaiges B, Borrós S, Pötgens AJ, Schmitz C, Sinner FM, Losen M, Steinbusch HW, Frank HG, Martinez-Martinez P, European NanoBioPharmaceutics Research Initiati - PLoS ONE (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, School for Mental Health and Neuroscience, Faculty of Health, Medicine and Life Sciences, Maastricht University, Maastricht, the Netherlands.

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.

No MeSH data available.


LC/MS chromatograms comparing different concentrations of peptide 5A in blood plasma matrix.Peptide 5A peaks in plasma matrix spiked with 0, 13.6 and 45.2 ng/mL (SRM trace: 877.7 → (412.2 + 341.1 + 946.4).
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pone.0126136.g004: LC/MS chromatograms comparing different concentrations of peptide 5A in blood plasma matrix.Peptide 5A peaks in plasma matrix spiked with 0, 13.6 and 45.2 ng/mL (SRM trace: 877.7 → (412.2 + 341.1 + 946.4).

Mentions: Peptide 5A used for generating the LC/MS standard curve was obtained by diluting the combined 5A+B peptide in plasma and subsequently reducing the disulfide bond between peptide 5A and peptide 5B. The LC/MS method showed good linearity in the range from 4.1 up to 1674 ng/mL (Fig 3A) with an expected lower limit of quantification of 13.5 ng/mL (S/N at 13.5 ng/mL was 330, Fig 4). Quality controls at 83.7 and 837 ng/mL measured in parallel with the in vivo study samples showed accuracies ranging between -7% and +7%.


Detection of Peptide-based nanoparticles in blood plasma by ELISA.

Bode GH, Pickl KE, Sanchez-Purrà M, Albaiges B, Borrós S, Pötgens AJ, Schmitz C, Sinner FM, Losen M, Steinbusch HW, Frank HG, Martinez-Martinez P, European NanoBioPharmaceutics Research Initiati - PLoS ONE (2015)

LC/MS chromatograms comparing different concentrations of peptide 5A in blood plasma matrix.Peptide 5A peaks in plasma matrix spiked with 0, 13.6 and 45.2 ng/mL (SRM trace: 877.7 → (412.2 + 341.1 + 946.4).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440766&req=5

pone.0126136.g004: LC/MS chromatograms comparing different concentrations of peptide 5A in blood plasma matrix.Peptide 5A peaks in plasma matrix spiked with 0, 13.6 and 45.2 ng/mL (SRM trace: 877.7 → (412.2 + 341.1 + 946.4).
Mentions: Peptide 5A used for generating the LC/MS standard curve was obtained by diluting the combined 5A+B peptide in plasma and subsequently reducing the disulfide bond between peptide 5A and peptide 5B. The LC/MS method showed good linearity in the range from 4.1 up to 1674 ng/mL (Fig 3A) with an expected lower limit of quantification of 13.5 ng/mL (S/N at 13.5 ng/mL was 330, Fig 4). Quality controls at 83.7 and 837 ng/mL measured in parallel with the in vivo study samples showed accuracies ranging between -7% and +7%.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, School for Mental Health and Neuroscience, Faculty of Health, Medicine and Life Sciences, Maastricht University, Maastricht, the Netherlands.

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.

No MeSH data available.