Labelling of live cells using fluorescent aptamers: Binding reversal with DNA nucleases

Hideyuki Terazono, Yu Anzai, Mikhail Soloviev, Kenji Yasuda

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A reversible cell labelling method has been developed for non-destructive and non-invasive cell labelling and purification. Our method uses high affinity single strand DNA (ssDNA) aptamers against surface exposed target molecules on cells. The aptamers are subsequently removed from the cell surface using DNase nuclease treatment. We exemplified our method by labelling human acute lymphoblastic leukemia cells with Qdot-ssDNA aptamers, and restoring them to the label-free condition by treatment with Benzonase. Binding of the fluorescent-aptamers to the cells was evaluated by measuring fluorescence intensity and was further confirmed using flow cytometry. Removal of the aptamers can be achieved in ~10 min by the DNase nuclease digestion. Incubation of cells with aptamers or with the nucleases results in no apparent damage to the cells and does not affect their growth rates. The latter were equivalent to the rates measured for the untreated cells. Our method provides an alternative to traditional antibody-based techniques and could be especially suitable for non-invasive reversible cell labelling and cell separations where maintaining native cell activity is needed.

Original languageEnglish
Article number8
JournalJournal of Nanobiotechnology
Volume8
DOIs
Publication statusPublished - 2010 Apr 13
Externally publishedYes

Fingerprint

Deoxyribonucleases
Labeling
Nucleotide Aptamers
Serratia marcescens nuclease
DNA
Flow cytometry
Antibodies
Purification
Labels
Fluorescence
Cells
Molecules
Cell Separation
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Digestion
Flow Cytometry

ASJC Scopus subject areas

  • Molecular Medicine
  • Bioengineering
  • Biomedical Engineering
  • Applied Microbiology and Biotechnology
  • Medicine (miscellaneous)
  • Pharmaceutical Science

Cite this

Labelling of live cells using fluorescent aptamers : Binding reversal with DNA nucleases. / Terazono, Hideyuki; Anzai, Yu; Soloviev, Mikhail; Yasuda, Kenji.

In: Journal of Nanobiotechnology, Vol. 8, 8, 13.04.2010.

Research output: Contribution to journalArticle

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