Orientation of capture antibodies on gold nanoparticles to improve the sensitivity of ELISA-based medical devices

Talanta. 2023 Aug 1:260:124650. doi: 10.1016/j.talanta.2023.124650. Epub 2023 May 5.

Abstract

The sensitivity of ELISA-based devices strongly depends on the right orientation of antibodies on the sensor surface. The aim of this work was to increase the analytical performance of a commercial ELISA-based medical device (VIDAS®), thanks to the specific orientation of antibodies on gold nanostructured disposables. For this purpose, fPSA VIDAS® assay was used as model and the disposable providing the antigen binding surface (SPR®) was functionalized with gold nanostructures coated with monovalent half-fragment antibodies (reduced IgG, rIgG). The functionalization of polystyrene SPRs® with gold nanostructures was achieved through a one-step incubation of gold dispersions in a mixture of non-toxic solvents. Five different concentrations of gold nanoparticles (NPs) were tested with a maximum fluorescence enhancement for NPs density around 3-8 *103 NPs/μm2 (752 ± 11 RFV vs 316 ± 5 RFV of bare SPRs®). The comparison of the dose-response curve obtained with commercial and gold coated-SPRs® revealed a significant improvement (p < 0.0001) of the analytical sensitivity of the VIDAS® system using nanostructured disposables. This improved version of SPRs® allows to distinguish small variations of fPSA concentrations opening the way to the application of this biomarker to other kinds of cancer as recently described in the literature.

Keywords: Analytical sensitivity; Controlled plastic decoration; Gold nanoparticles; Medical device; Oriented capture antibodies.

MeSH terms

  • Antibodies / chemistry
  • Biosensing Techniques*
  • Enzyme-Linked Immunosorbent Assay
  • Gold / chemistry
  • Metal Nanoparticles* / chemistry

Substances

  • Gold
  • Antibodies