Multiplexed protein detection by proximity ligation for cancer biomarker validation

Nat Methods. 2007 Apr;4(4):327-9. doi: 10.1038/nmeth1020. Epub 2007 Mar 18.

Abstract

We present a proximity ligation-based multiplexed protein detection procedure in which several selected proteins can be detected via unique nucleic-acid identifiers and subsequently quantified by real-time PCR. The assay requires a 1-microl sample, has low-femtomolar sensitivity as well as five-log linear range and allows for modular multiplexing without cross-reactivity. The procedure can use a single polyclonal antibody batch for each target protein, simplifying affinity-reagent creation for new biomarker candidates.

Publication types

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

MeSH terms

  • Antibodies / analysis*
  • Biomarkers, Tumor / analysis*
  • Neoplasm Proteins / analysis*
  • Neoplasms / diagnosis*
  • Neoplasms / metabolism
  • Oligonucleotides / analysis
  • Pilot Projects
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • Antibodies
  • Biomarkers, Tumor
  • Neoplasm Proteins
  • Oligonucleotides