A Facile Method to Quantify Synthetic Peptide Concentrations on Biomaterials

ACS Appl Mater Interfaces. 2024 Sep 18;16(37):49880-49888. doi: 10.1021/acsami.4c07164. Epub 2024 Sep 9.

Abstract

While it is well understood that peptides can greatly improve cell-material interactions, it is often challenging to determine the concentration of the peptide which decorates a material. Herein, we describe a straightforward method using readily, synthetically accessible Fmoc peptides and commercially available reagents to measure the concentration of peptides on nanoparticles, surfaces, and hydrogels. To achieve this, the Fmoc protecting group from immobilized peptides is removed under optimized basic conditions. The dibenzofulvene released can be quantified by HPLC or UV-vis spectroscopy, enabling a direct experimental measurement of the concentration of the peptide. We show that we can measure the concentration of a BMP-2 peptide mimic on a hydrogel to determine the concentration required to stimulate osteogenesis of human mesenchymal stem cells. We envision that this methodology will enable a more thorough understanding of the concentration of synthetic peptides decorated on many biomaterials (e.g., nanoparticles, surfaces, hydrogels) to improve deconvolution of the interactions at the cell-material interface.

Keywords: biomaterials; hydrogels; nanoparticles; peptides; quantification; surfaces.

MeSH terms

  • Biocompatible Materials* / chemical synthesis
  • Biocompatible Materials* / chemistry
  • Biocompatible Materials* / pharmacology
  • Bone Morphogenetic Protein 2* / chemistry
  • Humans
  • Hydrogels* / chemical synthesis
  • Hydrogels* / chemistry
  • Hydrogels* / pharmacology
  • Mesenchymal Stem Cells* / cytology
  • Mesenchymal Stem Cells* / drug effects
  • Nanoparticles / chemistry
  • Osteogenesis / drug effects
  • Peptides* / chemistry
  • Peptides* / pharmacology

Substances

  • Peptides
  • Hydrogels
  • Bone Morphogenetic Protein 2
  • Biocompatible Materials
  • BMP2 protein, human