Development of an Enzyme-Linked ImmunoSorbent Assay (ELISA) with Enhanced Sensitivity in a Nanofluidic System

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.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Electrical Engineering and Computer Science.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2009

Author dc:creator

• Cheow, Lih Feng

Advisor dc:contributor.advisor

• Jongyoon Han.

• Electrical Engineering and Computer Science.

Statement dc:rights

• 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.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/55148

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/55148