Preparation of fractioned DNA aptamer-Pt complex through ultrafiltration and the colorimetric sensing of thrombin

Akon Higuchi, Siou Ting Yang, Yi Di Siao, Pei Vin Hsieh, Hisashi Fukushima, Yung Chang, Wen Yih Chen

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

DNA aptamers were combined with platinum complexes to form DNA-Pt complexes, which exhibited peroxidase enzymatic activity while retaining the specific binding ability of aptamers. The DNA-Pt complexes were fractioned by size through ultrafiltration membranes having several molecular weight cut-offs, and the peroxidase enzymatic activities of these fractions were compared. The enzymatic activity was highest in the filtrate of DNA-Pt complex solutions prepared with cisplatin and K2[PtCl4] after ultrafiltration through membranes having MWCO = 300,000. These showed 1.2-fold and 1.6-fold higher activity, respectively, than the corresponding unfractioned complexes. Sandwich-type DNAzyme-linked aptamer assays (DLAAs) using unfractioned or fractioned DNA-Pt complexes successfully detected the target protein thrombin. DLAAs incorporating a DNA-Pt (K2[PtCl4]) complex fractioned through ultrafiltration membranes having MWCO = 300,000 showed the highest sensitivity among DLAAs made using fractioned DNA-Pt complexes, and had a 13-fold higher sensitivity than those made with unfractioned DNA-Pt complexes.

Original languageEnglish
Pages (from-to)97-103
Number of pages7
JournalJournal of Membrane Science
Volume328
Issue number1-2
DOIs
StatePublished - 5 Feb 2009

Keywords

  • Aptamer
  • Colorimetric sensing
  • DNA
  • DNAzyme
  • Platinum complex
  • Ultrafiltration

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