Stable and scalable SERS tags conjugated with neutravidin for the detection of fibroblast activation protein (FAP) in primary fibroblasts

Nanotechnology. 2021 Apr 30;32(29). doi: 10.1088/1361-6528/abf5fd.

Abstract

SERS tags are a class of nanoparticles with great potential in advanced imaging experiments. The preparation of SERS tags however is complex, as they suffer from the high variability of the SERS signals observed even at the slightest sign of aggregation. Here, we developed a method for the preparation of SERS tags based on the use of gold nanostars conjugated with neutravidin. The SERS tags here obtained are extremely stable in all biological buffers commonly employed and can be prepared at a relatively large scale in very mild conditions. The obtained SERS tags have been used to monitor the expression of fibroblast activation protein alpha (FAP) on the membrane of primary fibroblasts obtained from patients affected by Crohn's disease. The SERS tags allowed the unambiguous identification of FAP on the surface of cells thus suggesting the feasibility of semi-quantitative analysis of the target protein. Moreover, the use of the neutravidin-biotin system allows to apply the SERS tags for any other marker detection, for example, different cancer cell types, simply by changing the biotinylated antibody chosen in the analysis.

Keywords: SERS tags; fibroblast activation protein; gold nanostars; neutravidin; plasmonics.

MeSH terms

  • Avidin / chemistry
  • Biotin / chemistry
  • Crohn Disease / genetics
  • Crohn Disease / metabolism
  • Crohn Disease / pathology
  • Endopeptidases / analysis
  • Endopeptidases / genetics*
  • Endopeptidases / metabolism
  • Gene Expression
  • Gold / chemistry
  • Humans
  • Ileum / metabolism
  • Ileum / pathology
  • Membrane Proteins / analysis
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Metal Nanoparticles / chemistry*
  • Metal Nanoparticles / ultrastructure
  • Myofibroblasts / metabolism*
  • Myofibroblasts / pathology
  • Octoxynol / chemistry*
  • Polyethylene Glycols / chemistry
  • Primary Cell Culture
  • Spectrum Analysis, Raman / methods*
  • Staining and Labeling

Substances

  • Membrane Proteins
  • neutravidin
  • Avidin
  • Polyethylene Glycols
  • Biotin
  • Gold
  • Octoxynol
  • Endopeptidases
  • fibroblast activation protein alpha