Chemiluminescence detection of a protein through the aptamer-controlled catalysis of a porphyrin probe

Anal Chem. 2015 Aug 18;87(16):8336-41. doi: 10.1021/acs.analchem.5b01511. Epub 2015 Aug 6.

Abstract

Sensitive and selective protein detection based on the aptamer-controlled noncovalent porphyrin probe self-assembly is reported for the first time. Vascular endothelial growth factor (VEGF) is a predominant biomarker in cancer angiogenesis. In this work, a positively charged porphyrin probe, manganese(III) meso-tetrakis(N-methylpyridinum-4-yl)porphyrin (Mn-PyP), was prepared. Using it as a catalyst, a label-free chemiluminescence (CL) turn-on approach for sensitive VEGF detection is developed. Mn-PyP could catalyze the luminol CL reaction. The VEGF aptamer could induce aggregation of Mn-PyP. As a result, the Mn-PyP-catalyzed CL reaction is efficiently suppressed. Upon the addition of VEGF, the specific binding of VEGF to the aptamer weakens the interactions between the aptamer and Mn-PyP. The Mn-PyP monomers are released, and a turn-on CL signal is thus detected. Our method is quite sensitive; 50 pM of VEGF could be easily detected. It is also very selective against other proteins. Our assay provides an aptamer-based efficient way for protein quantification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Catalysis
  • Chemistry Techniques, Analytical / methods*
  • Fluorescent Dyes / metabolism
  • Limit of Detection
  • Luminescence*
  • Manganese / chemistry*
  • Molecular Structure
  • Porphyrins / chemistry
  • Porphyrins / metabolism*
  • Proteins / analysis*
  • Proteins / chemistry
  • Vascular Endothelial Growth Factor A / analysis
  • Vascular Endothelial Growth Factor A / chemistry

Substances

  • Aptamers, Nucleotide
  • Fluorescent Dyes
  • Porphyrins
  • Proteins
  • Vascular Endothelial Growth Factor A
  • Manganese