PT  - JOURNAL ARTICLE
AU  - Sakamoto, Shingo
AU  - Komatsu, Toru
AU  - Watanabe, Rikiya
AU  - Zhang, Yi
AU  - Inoue, Taiki
AU  - Kawaguchi, Mitsuyasu
AU  - Nakagawa, Hidehiko
AU  - Ueno, Takaaki
AU  - Okusaka, Takuji
AU  - Honda, Kazufumi
AU  - Noji, Hiroyuki
AU  - Urano, Yasuteru
TI  - Multiplexed single-molecule enzyme activity analysis for counting disease-related proteins in biological samples
AID  - 10.1126/sciadv.aay0888
DP  - 2020 Mar 01
TA  - Science Advances
PG  - eaay0888
VI  - 6
IP  - 11
4099  - http://advances.sciencemag.org/content/6/11/eaay0888.short
4100  - http://advances.sciencemag.org/content/6/11/eaay0888.full
SO  - Sci Adv2020 Mar 01; 6
AB  - 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.