Digital concentration readout of single enzyme molecules using femtoliter arrays and Poisson statistics

Nano Letters
David M Rissin, David R Walt

Abstract

Methods for accurately quantifying the concentration of a particular analyte in solution are all based on ensemble responses in which many analyte molecules give rise to the measured signal. In this paper, single molecules of beta-galactosidase were monitored using a 1 mm diameter fiber optic bundle with 2.4 x 10(5) individually sealed, femtoliter microwell reactors. By observation of the buildup of fluorescent products from single enzyme molecule catalysis over the array of reaction vessels and by application of a Poisson statistical analysis, a digital concentration readout was obtained. This approach should prove useful for single molecule enzymology and ultrasensitive bioassays. More generally, the ability to determine concentration by counting individual molecules offers a new approach to analysis of dilute solutions.

References

Oct 27, 2001·Biosensors & Bioelectronics·H NagaiE Tamiya
Apr 17, 2003·Journal of Biotechnology·Michihiko NakanoAkira Mizuno
Jul 10, 2003·Proceedings of the National Academy of Sciences of the United States of America·Dhaval N Gosalia, Scott L Diamond
Aug 30, 2003·Science·Everett A LipmanWilliam A Eaton
Oct 2, 2003·Nature Biotechnology·George M Whitesides
Oct 23, 2003·Analytical Chemistry·Aaron R WheelerAntoine Daridon
Mar 17, 2004·Analytical Chemistry·Mathieu FoquetHarold G Craighead
Feb 22, 2005·Nature Biotechnology·Yannick RondelezHiroyuki Noji

❮ Previous
Next ❯

Citations

Nov 17, 2009·Analytical and Bioanalytical Chemistry·F DeissP R Stoddart
Feb 9, 2012·Lab on a Chip·Huaibin ZhangDavid R Walt
Jul 31, 2013·Analytical and Bioanalytical Chemistry·Venkata Suresh VajralaStéphane Arbault
Jan 15, 2014·Omics : a Journal of Integrative Biology·Firas H KobeissyKevin Wang
Oct 30, 2007·Proceedings of the National Academy of Sciences of the United States of America·Hans H GorrisDavid R Walt
May 16, 2014·Lab on a Chip·David R Walt
Sep 4, 2014·Lab on a Chip·Baichen LiZhenyu Li
Oct 30, 2010·Lab on a Chip·Shouichi SakakiharaHiroyuki Noji
Sep 11, 2012·Lab on a Chip·Soo Hyeon KimHiroyuki Noji
Jun 21, 2013·The Analyst·Soo Hyeon KimDominique Fourmy
Aug 20, 2009·Chemical Society Reviews·Qi ChenG Julius Vancso
Oct 5, 2013·Chemical Society Reviews·Jon R AskimKenneth S Suslick
Sep 16, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Raphaela B Liebherr, Hans H Gorris
Oct 17, 2007·The FEBS Journal·Hans H GorrisDavid R Walt
Jul 13, 2007·Molecular Systems Biology·Mitchel J Doktycz, Michael L Simpson
Aug 9, 2015·Analytical and Bioanalytical Chemistry·Raphaela B LiebherrHans H Gorris
Apr 22, 2015·Trends in Biotechnology·Marina CretichMarcella Chiari
Apr 3, 2009·Journal of the American Chemical Society·Hans H Gorris, David R Walt
Jul 30, 2014·Analytical Chemistry·Matteo CornagliaMartin A M Gijs
Oct 23, 2016·Analytica Chimica Acta·Elena Benito-PeñaMaria C Moreno-Bondi
Mar 8, 2017·ACS Applied Materials & Interfaces·Deborah DecropJeroen Lammertyn
Mar 1, 2018·Lab on a Chip·Lei BaoXuehua Zhang
Dec 5, 2006·BioTechniques·David R Walt
Jul 23, 2016·Angewandte Chemie·J Justin Gooding, Katharina Gaus
Aug 21, 2018·Lab on a Chip·Andreas H KundingMartin Dufva
Sep 13, 2008·Chembiochem : a European Journal of Chemical Biology·Hiroshi KitaTetsuya Yomo
Apr 27, 2010·Angewandte Chemie·Hans H Gorris, David R Walt
Jan 30, 2019·Clinical Chemistry·David C Duffy, David R Walt
Sep 21, 2018·BMJ Open Sport & Exercise Medicine·Colin WallacePaul van Donkelaar

❮ Previous
Next ❯

Related Concepts

Related Feeds

Researcher Network:CZI Neurodegeneration Challenge

The Neurodegeneration Challenge Network aims to provide funding for and to bring together researchers studying neurodegenerative diseases. Find the latest research from the NDCN grantees here.