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Analytical and Bioanalytical Chemistry

, Volume 408, Issue 3, pp 875–884 | Cite as

β-Conglutin dual aptamers binding distinct aptatopes

  • Miriam Jauset Rubio
  • Markéta Svobodová
  • Teresa Mairal
  • Thomas Schubert
  • Stefan Künne
  • Günter Mayer
  • Ciara K. O’SullivanEmail author
Research Paper

Abstract

An aptamer was previously selected against the anaphylactic allergen β-conglutin (β-CBA I), which was subsequently truncated to an 11-mer and the affinity improved by two orders of magnitude. The work reported here details the selection and characterisation of a second aptamer (β-CBA II) selected against a second aptatope on the β-conglutin target. The affinity of this second aptamer was similar to that of the 11-mer, and its affinity was confirmed by three different techniques at three independent laboratories. This β-CBA II aptamer in combination with the previously selected β-CBA I was then exploited to a dual-aptamer approach. The specific and simultaneous binding of the dual aptamer (β-CBA I and β-CBA II) to different sites of β-conglutin was confirmed using both microscale thermophoresis and surface plasmon resonance where β-CBA II serves as the primary capturing aptamer and β-CBA I or the truncated β-CBA I (11-mer) as the secondary signalling aptamer, which can be further exploited in enzyme-linked aptamer assays and aptasensors.

Keywords

β-Conglutin Dual aptamer SELEX Sandwich assay Truncation studies 

Notes

Acknowledgments

This work was supported by funding from the national project RecerCaixa (CO074670 APTALUP).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

216_2015_9179_MOESM1_ESM.pdf (1.9 mb)
ESM 1 (PDF 1971 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Miriam Jauset Rubio
    • 1
  • Markéta Svobodová
    • 1
  • Teresa Mairal
    • 1
  • Thomas Schubert
    • 2
  • Stefan Künne
    • 3
  • Günter Mayer
    • 3
  • Ciara K. O’Sullivan
    • 1
    • 4
    Email author
  1. 1.Nanobiotechnology and Bioanalysis Group, Department of Chemical EngineeringUniversitat Rovira I VirgiliTarragonaSpain
  2. 2.2bind GmbHRegensburgGermany
  3. 3.Life and Medical Sciences InstituteUniversity of BonnBonnGermany
  4. 4.Institució Catalana de Recerca I Estudis AvancatsBarcelonaSpain

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