Protein biomarker enrichment by biomarker antibody complex elution for immunoassay biosensing

Biosens Bioelectron. 2011 Nov 15;29(1):18-22. doi: 10.1016/j.bios.2011.06.046. Epub 2011 Jul 21.

Abstract

It is very challenging to perform sample enrichment for protein biomarkers because proteins can easily change conformation and denature. In this paper we demonstrate protein enrichment suited for high-sensitivity integrated immuno-biosensing. The method enhances the concentration of the biomarkers and simultaneously removes matrix components that could interfere with the immunoassay. Biomarkers are captured using antibody coated magnetic particles and the biomarker antibody complexes are released by enzymatic elution. The eluted complexes are subsequently detected in a sandwich immunoassay biosensor. A scaling study of the enrichment process demonstrates an enrichment factor of 15 in buffer and plasma. We analyze the enrichment factor in terms of the three basic steps of the assay (capture, concentration, elution) and we quantify their respective efficiencies. The process is suited for integration into bio-analytical tools.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen-Antibody Complex / isolation & purification
  • Biomarkers / analysis
  • Biomarkers, Tumor / blood
  • Biosensing Techniques / methods*
  • Epitopes / isolation & purification
  • Humans
  • Immunoassay / methods*
  • Magnetics
  • Male
  • Prostate-Specific Antigen / blood
  • Prostatic Neoplasms / blood
  • Prostatic Neoplasms / diagnosis
  • Proteins / immunology
  • Proteins / isolation & purification*

Substances

  • Antigen-Antibody Complex
  • Biomarkers
  • Biomarkers, Tumor
  • Epitopes
  • Proteins
  • Prostate-Specific Antigen