Mikhail Soloviev, Hideyuki Terazono, Yu Anzai and Kenji Yasuda (2010) Labelling of live cells using fluorescent aptamers: binding reversal with DNA nucleases. Journal of Nanobiotechnology, 8 (8). pp. . ISSN 1477-3155
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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.
This is a Published version This version's date is: 13/04/2010 This item is peer reviewed
https://repository.royalholloway.ac.uk/items/58b683b6-0fc0-628d-ad0a-a4ef18cbd1bc/1/
Deposited by () on 08-Mar-2011 in Royal Holloway Research Online.Last modified on 08-Mar-2011
© 2010 Terazono et al; licensee BioMed Central Ltd.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.