Quantitative label-free screening for antibodies using scattering biophotonic microarray imaging

Anal Biochem. 2010 Jan 1;396(1):30-5. doi: 10.1016/j.ab.2009.08.008. Epub 2009 Aug 11.

Abstract

A biophotonic array based on gold nanoparticles functionalized with antigen proteins has been used to determine the concentrations of the respective antibodies in solution. Four proteins-fibrinogen, bovine serum albumin, transferrin, and C-reactive protein-were used to construct a test array with the assay repeated a number of times. The antibody-antigen association and dissociation rate constants were determined for the antibody assays from a series of calibration experiments. The label-free determination of the unknown antibody concentrations was performed using two related kinetic analyses. From these results, the current array assay sensitivity is 250 ng ml(-1) with an accuracy of 15% using an 8-min kinetic measurement and a 16-spot averaged assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / analysis*
  • Antibodies / immunology
  • Antigens / immunology
  • Biosensing Techniques
  • Cattle
  • High-Throughput Screening Assays / methods*
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Kinetics
  • Microarray Analysis / methods*
  • Photons*
  • Staining and Labeling*

Substances

  • Antibodies
  • Antigens