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Investigation and verification of a bioluminescent biosensor for the quantitation of ara-CTP generation: A biomarker for cytosine arabinoside sensitivity in acute myeloid leukaemia

Graham Smith, J.; Peter Fitzgerald, S.; Martin, Ashley; Ann Smith, M.; Conway, Myra; Anderson, Elizabeth; Conway, Myra E.; Alloush, Habib; O'Malley, Kieran; Smith, Ann; Ashley, Martinc; Ruddock, Mark; Reid, Cherith; Lamont, John; Fitzgerald, Peter; Smith, Graham; Mehta, Priyanka; Salisbury, Vyv

Authors

J. Graham Smith

S. Peter Fitzgerald

Ashley Martin

M. Ann Smith

Myra Conway

Myra Conway Myra.Conway@uwe.ac.uk
Occasional Associate Lecturer - CHSS - DAS

Habib Alloush

Kieran O'Malley

Ann Smith

Martinc Ashley

Mark Ruddock

Cherith Reid

John Lamont

Peter Fitzgerald

Graham Smith

Priyanka Mehta

Vyv Salisbury



Abstract

A novel whole cell bacterial biosensor, which emits light in response to the active metabolite of cytosine arabinoside (ara-C, cytarabine), ara-CTP, has been investigated and verified. The biosensor has been formulated as an ex vivo assay, designed for peripheral blood or bone marrow cells, which can produce a clinical result within a working day. The nucleoside analogue ara-C is a key agent for treatment of acute myeloid leukaemia (AML); treatment decisions are made rapidly with AML, patients often receiving same-day commencement of chemotherapy. Currently no rapid predictive test is available to select appropriate therapy for patients prior to treatment. Experiments were designed to determine optimal assay conditions using leukaemic cell lines. We observed a significant increase (~15 fold) in bioluminescence signal compared to control after 8-h incubation of the biosensor with ara-C. This corresponded to a >2-log increase in light output per bacterial cell. Interestingly, bioluminescence conferred a survival advantage to the bacteria following ara-C treatment. The assay is sensitive (lower limit of quantitation of 0.05. μM), selective, accurate (≤15% RE) and precise (≤15% coefficient of variation) over a linear concentration range of ara-CTP (0.05-0.5. μM), and detection is independent of reaction volume. Recovery of added standard was tested using ex vivo patient leukaemic cells (n=5). Stability studies on lyophilized bacterial biosensor were performed to ensure maintenance of performance over 12 months. The biosensor assay could be invaluable to the clinician, assisting with treatment selection, and potentially mitigating the risks of resistance and toxicity observed with this drug. © 2013 Elsevier B.V.

Journal Article Type Article
Acceptance Date Sep 6, 2013
Publication Date Feb 15, 2014
Journal Biosensors and Bioelectronics
Print ISSN 0956-5663
Electronic ISSN 1873-4235
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 52
Pages 345-353
DOI https://doi.org/10.1016/j.bios.2013.09.014
Keywords whole cell biosensor, bioluminescent, leukaemia, chemotherapy
Public URL https://uwe-repository.worktribe.com/output/821121
Publisher URL http://dx.doi.org/10.1016/j.bios.2013.09.014