Minimizing antibody cross-reactivity in multiplex detection of biomarkers in paper-based point-of-care assays

Nanoscale. 2016 Apr 21;8(15):8195-201. doi: 10.1039/c5nr09207h.

Abstract

Highly multiplexed immunoassays could allow convenient screening of hundreds or thousands of protein biomarkers simultaneously in a clinical sample such as serum or plasma, potentially allowing improved diagnostic accuracy and clinical management of many conditions such as autoimmune disorders, infections, and several cancers. Currently, antibody microarray-based tests are limited in part due to cross reactivity from detection antibody reagents. Here we present a strategy that reduces the cross-reactivity between nanoparticle-bound reporter antibodies through the application of ultrasound energy. By this concept, it was possible to achieve a sensitivity 10(3)-fold (5 pg mL(-1)) lower than when no ultrasound was applied (50 ng mL(-1)) for the simultaneous detection of three different antigens. The detection limits and variability achieved with this technique rival those obtained with other types of multiplex sandwich assays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Immobilized / immunology*
  • Biomarkers / blood*
  • Cross Reactions
  • Gold
  • Humans
  • Immunoassay / methods*
  • Immunoassay / statistics & numerical data
  • Limit of Detection
  • Metal Nanoparticles
  • Point-of-Care Systems* / statistics & numerical data
  • Sensitivity and Specificity
  • Ultrasonics

Substances

  • Antibodies, Immobilized
  • Biomarkers
  • Gold