Label-free and washing-free alkaline phosphatase assay using a personal glucose meter

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We herein describe a personal glucose meter (PGM)-based method for a label-free and washing-free determination of alkaline phosphatase (ALP) activity, which relies on the cascade enzymatic reactions promoted by hexokinase and pyruvate kinase to couple ALP activity with the amount of glucose. In principle, the presence of target ALP scavenges on adenosine 5'-triphosphate (ATP), a phosphate source for hexokinase-catalyzed reactions, and thus suppresses the ensuing cascade enzymatic reactions. As a result, the initial high amount of glucose is maintained and the amount of glucose, which is proportional to ALP activity, is simply measured by a hand-held PGM. Based on this novel strategy, we successfully determined the ALP activity down to 8.9 U/L with the high selectivity. In addition, the diagnostic capability of this method was demonstrated by reliably assaying the ALP activity in non-diluted human blood without any pretreatment steps.
Publisher
BMC
Issue Date
2019-06
Language
English
Article Type
Letter
Citation

JOURNAL OF BIOLOGICAL ENGINEERING, v.13

ISSN
1754-1611
DOI
10.1186/s13036-019-0182-3
URI
http://hdl.handle.net/10203/263242
Appears in Collection
CBE-Journal Papers(저널논문)
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