Journal
SCIENCE ADVANCES
Volume 6, Issue 11, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/sciadv.aay0888
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Funding
- MEXT [24655147, 15H05371, 15 K14937, 17 K19477, 18H04538, 19H02846, 15 K18899, 16H05103]
- JST (PRESTO)
- JST (PRESTO Network)
- Council for Science, Technology, and Innovation (Cabinet office, Government of Japan)
- AMED [RRIME]
- JSPS
- Naito Foundation
- Mochida Memorial Foundation for Medical and Pharmaceutical Research
- Tokyo Biochemical Research Foundation
- AMED [CREST]
- AMED [P-CREATE] [18cm0106403h0003]
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We established an ultrasensitive method for identifying multiple enzymes in biological samples by using a multiplexed microdevice-based single-molecule enzymatic assay. We used a paradigm in which we count the number of enzyme molecules by profiling their single enzyme activity characteristics toward multiple substrates. In this proof-of-concept study of the single enzyme activity-based protein profiling (SEAP), we were able to detect the activities of various phosphoric ester-hydrolyzing enzymes such as alkaline phosphatases, tyrosine phosphatases, and ectonucleotide pyrophosphatases in blood samples at the single-molecule level and in a subtype-discriminating manner, demonstrating its potential usefulness for the diagnosis of diseases based on ultrasensitive detection of enzymes.
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