Multiplexed cancer biomarker detection using quartz-based photonic crystal surfaces

Anal Chem. 2012 Jan 17;84(2):1126-33. doi: 10.1021/ac202817q. Epub 2011 Dec 29.

Abstract

A photonic crystal (PC) surface is demonstrated as a high-sensitivity platform for detection of a panel of 21 cancer biomarker antigens using a sandwich enzyme-linked immunosorbent assay (ELISA) microarray format. A quartz-based PC structure fabricated by nanoimprint lithography, selected for its low autofluorescence, supports two independent optical resonances that simultaneously enable enhancement of fluorescence detection of biomarkers and label-free quantification of the density of antibody capture spots. A detection instrument is demonstrated that supports fluorescence and label-free imaging modalities, with the ability to optimize the fluorescence enhancement factor on a pixel-by-pixel basis throughout the microarray using an angle-scanning approach for the excitation laser that automatically compensates for variability in surface chemistry density and capture spot density. Measurements show that the angle-scanning illumination approach reduces the coefficient of variation of replicate assays by 20-99% compared to ordinary fluorescence microscopy, thus supporting reduction in limits of detectable biomarker concentration. Using the PC resonance, biomarkers in mixed samples were detectable at the lowest concentrations tested (2.1-41 pg/mL), resulting in a three-log range of quantitative detection.

Publication types

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

MeSH terms

  • Antibodies, Immobilized / chemistry*
  • Antibodies, Immobilized / metabolism
  • Biomarkers, Tumor / analysis*
  • Biosensing Techniques*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunoassay
  • Lasers
  • Neoplasms / immunology
  • Neoplasms / metabolism*
  • Photons*
  • Protein Array Analysis
  • Quartz*
  • Spectrometry, Fluorescence
  • Surface Properties

Substances

  • Antibodies, Immobilized
  • Biomarkers, Tumor
  • Quartz