RT Journal Article
SR Electronic
T1 Multiplexed single-molecule enzyme activity analysis for counting disease-related proteins in biological samples
JF Science Advances
JO Sci Adv
FD American Association for the Advancement of Science
SP eaay0888
DO 10.1126/sciadv.aay0888
VO 6
IS 11
A1 Sakamoto, Shingo
A1 Komatsu, Toru
A1 Watanabe, Rikiya
A1 Zhang, Yi
A1 Inoue, Taiki
A1 Kawaguchi, Mitsuyasu
A1 Nakagawa, Hidehiko
A1 Ueno, Takaaki
A1 Okusaka, Takuji
A1 Honda, Kazufumi
A1 Noji, Hiroyuki
A1 Urano, Yasuteru
YR 2020
UL http://advances.sciencemag.org/content/6/11/eaay0888.abstract
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.