Application of the Open qPCR Instrument for the in Vitro Selection of DNA Aptamers against Epidermal Growth Factor Receptor and Drosophila C Virus

Tulsi Ram Damase, Tanya A Miura, Christine E Parent, Peter B Allen

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

The low-cost Open qPCR instrument can be used for different tasks in the aptamer selection process: quantification of DNA, cycle course optimization, screening, and final binding characterization. We have selected aptamers against whole Drosophila C virus (DCV) particles and recombinant epidermal growth factor receptor (EGFR). We performed systematic evolution of ligands by exponential enrichment (SELEX) using the Open qPCR to optimize each amplification step. The Open qPCR instrument identified the best aptamer candidate. The Open qPCR has the capacity to perform melt curves, and we used this function to perform thermofluorimetric analysis (TFA) to quantify target-aptamer binding. We confirmed target-aptamer binding using flow cytometry. A sandwich type luminescence bioassay based on our anti-DCV aptamer was sensitive to DCV and did not respond to a related virus, demonstrating that our selected anti-DCV aptamer can be used to specifically detect DCV.

Original languageEnglish (US)
Pages (from-to)45-54
Number of pages10
JournalACS combinatorial science
Volume20
Issue number2
DOIs
StatePublished - Feb 12 2018

Keywords

  • Aptamers, Nucleotide/chemistry
  • Binding, Competitive
  • Combinatorial Chemistry Techniques
  • Dicistroviridae/isolation & purification
  • ErbB Receptors/antagonists & inhibitors
  • Fluorescence
  • Gene Library
  • Ligands
  • Microspheres
  • Polymerase Chain Reaction/methods
  • SELEX Aptamer Technique

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