Sub-zeptomole Detection of Biomarker Proteins Using a Microfluidic Immunoarray with Nanostructured Sensors

Anal Chem. 2020 Jun 16;92(12):8021-8025. doi: 10.1021/acs.analchem.0c01507. Epub 2020 May 27.

Abstract

We report here a low-cost electrochemical immunoarray with unprecedented sensitivity in the sub-zeptomole range with up to 5 log-decades dynamic range for accurate, multiplexed protein determinations. The microfluidic array features eight carbon sensors coated with a dense layer of 5 nm gold-nanoparticles derivatized with primary antibodies. Analyte proteins are captured by secondary antibody-poly-HPR (horseradish peroxidase) bioconjugates containing 400 HRP enzyme labels, with amplified amperometric peaks developed using H2O2 activator and hydroquinone mediator. Prostate cancer biomarkers prostate specific antigen (PSA), vascular endothelial growth factor-D (VEGF-D), ETS-related gene protein (ERG), and insulin-like growth factor-1 (IGF-1) were measured simultaneously with sub-fg/mL LODs (0.08-0.22 zmol). These proteins were determined in serum of postprostatectomy cancer patients which had much lower levels than prostate cancer patients without surgery. This immunoassay protocol makes thousands of low-abundance proteins accessible to quantitative measurements down to zeptomole levels.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / analysis*
  • Biosensing Techniques*
  • Electrochemical Techniques*
  • Humans
  • Immunoassay*
  • Insulin-Like Growth Factor I / analysis
  • Lab-On-A-Chip Devices*
  • Male
  • Nanostructures / chemistry*
  • Prostate-Specific Antigen / analysis
  • Prostatic Neoplasms / diagnosis*
  • Transcriptional Regulator ERG / analysis
  • Vascular Endothelial Growth Factor D / analysis

Substances

  • Biomarkers, Tumor
  • ERG protein, human
  • IGF1 protein, human
  • Transcriptional Regulator ERG
  • VEGFD protein, human
  • Vascular Endothelial Growth Factor D
  • Insulin-Like Growth Factor I
  • Prostate-Specific Antigen