Terminal protection of small molecule-linked ssDNA for label-free and sensitive fluorescent detection of folate receptor

Talanta. 2014 Oct:128:237-41. doi: 10.1016/j.talanta.2014.05.009. Epub 2014 May 13.

Abstract

In this work, based on terminal protection of folate-linked ssDNA (FA-ssDNA) and the SYBR Gold fluorescent dye, we describe the development of a label-free fluorescent strategy for the detection of folate receptors (FRs). The binding between the target FR and the FA moiety of the FA-ssDNA protects the FR bound FA-ssDNA from digesting by Exo I. The binding of SYBR Gold to the terminal protected, un-digested FA-ssDNA leads to enhanced fluorescent emission for the monitoring of FR with a detection limit of 30 pM. Besides, the developed method also shows high selectivity toward FR against other control proteins. Moreover, our approach avoids the labeling of the probes with fluorescent tags and achieves label-free detection of FR. With these advantages, the proposed method thus holds promising potential for the development of simple and convenient strategies for the detection of other proteins by using different small molecule receptor/protein ligand pairs.

Keywords: Fluorescence; Folate receptors; Folate-linked ssDNA; Terminal protection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Single-Stranded / chemistry*
  • Fluorescent Dyes / chemistry*
  • Folate Receptors, GPI-Anchored / analysis*
  • Folate Receptors, GPI-Anchored / blood
  • Folate Receptors, GPI-Anchored / chemistry
  • Folic Acid / chemistry*
  • Humans
  • Organic Chemicals / chemistry
  • Reproducibility of Results

Substances

  • DNA, Single-Stranded
  • Fluorescent Dyes
  • Folate Receptors, GPI-Anchored
  • Organic Chemicals
  • SYBR Gold nucleic acid gel stain
  • Folic Acid