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Jongyoon Han. Cheow, Lih Feng Massachusetts Institute of Technology. Dept. of Electrical Engineering and Computer Science. Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science 2010-05-25T20:53:59Z 2010-05-25T20:53:59Z 2009 2009 http://hdl.handle.net/1721.1/55148 599993015 Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2009. Cataloged from PDF version of thesis. Includes bibliographical references (p. 66-68). Experimental studies were performed to evaluate the kinetics and equilibrium binding constants of biomolecules in nanofluidic channels. Binding events in the nanochannel were detected using electrical and fluorescence methods. We concluded that antibody-antigen binding constants in nanochannels were similar to experiments performed in microtiter plates at low antigen concentrations; however the bound fraction in nanochannels at high antigen concentration decreased due to steric hindrance. Binding kinetics in nanochannels was limited by convective transport of analytes, instead of diffusion or reaction. We also found that enzymatic reactions in nanochannels were very effective due to short diffusion length and high surface area to volume ratio. A bead based ELISA was developed to exploit the rapid binding reactions in the bulk and efficient enzymatic conversion in the nanochannels. Additionally, electrokinetic concentrators were integrated with multiplexed bead based ELISA to further improve the detection sensitivity of a sandwich immunoassay. by Lih Feng Cheow. S.M. 68 p. eng Massachusetts Institute of Technology M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 Electrical Engineering and Computer Science. Development of an Enzyme-Linked ImmunoSorbent Assay (ELISA) with Enhanced Sensitivity in a Nanofluidic System Development of an Enzyme-Linked Immuno Sorbent Assay (ELISA) with Enhanced Sensitivity in a Nanofluidic System Development of an ELISA with Enhanced Sensitivity in a Nanofluidic System Thesis application/pdf false Publication unknown unknown <Publication xmlns="https://www.openaire.eu/cerif-profile/1.1/" id="f3805646-3895-4d54-bb23-0defe22d90d3"> <Type xmlns="https://www.openaire.eu/cerif-profile/vocab/COAR_Publication_Types">http://purl.org/coar/resource_type/c_1843</Type> <Language>eng</Language> <Title>Development of an Enzyme-Linked ImmunoSorbent Assay (ELISA) with Enhanced Sensitivity in a Nanofluidic System</Title> <Subtitle>Development of an Enzyme-Linked Immuno Sorbent Assay (ELISA) with Enhanced Sensitivity in a Nanofluidic System</Subtitle> <Subtitle>Development of an ELISA with Enhanced Sensitivity in a Nanofluidic System</Subtitle> <PublishedIn> <Publication> </Publication> </PublishedIn> <PublicationDate>2009</PublicationDate> <Authors> <Author> <DisplayName>Cheow, Lih Feng</DisplayName> <Affiliation> <OrgUnit> </OrgUnit> </Affiliation> </Author> </Authors> <Editors> </Editors> <Publishers> <Publisher> <DisplayName>Massachusetts Institute of Technology</DisplayName> <OrgUnit /> </Publisher> </Publishers> <License>http://dspace.mit.edu/handle/1721.1/7582</License> <Keyword>Electrical Engineering and Computer Science.</Keyword> <Abstract>Experimental studies were performed to evaluate the kinetics and equilibrium binding constants of biomolecules in nanofluidic channels. Binding events in the nanochannel were detected using electrical and fluorescence methods. We concluded that antibody-antigen binding constants in nanochannels were similar to experiments performed in microtiter plates at low antigen concentrations; however the bound fraction in nanochannels at high antigen concentration decreased due to steric hindrance. Binding kinetics in nanochannels was limited by convective transport of analytes, instead of diffusion or reaction. We also found that enzymatic reactions in nanochannels were very effective due to short diffusion length and high surface area to volume ratio. A bead based ELISA was developed to exploit the rapid binding reactions in the bulk and efficient enzymatic conversion in the nanochannels. Additionally, electrokinetic concentrators were integrated with multiplexed bead based ELISA to further improve the detection sensitivity of a sandwich immunoassay.</Abstract> <Access xmlns="http://purl.org/coar/access_right" > </Access> </Publication>