Rapid detection of two-protein interaction with a single fluorophore by using a microfluidic device. Academic Article

abstract

• We have developed a microfluidics based platform and methodology named MAPS (microfluidic system for analyzing proteins in single complex) for detecting two protein interactions rapidly using a single fluorophore. Target proteins were labelled with Quantum dot 525 (QD525) via specific polyclonal antibodies, and were transported through the microfluidic channel subsequently, where the 375 nm excitation laser light was focused to form a detection volume. Photon bursts from target proteins passing through the detection volume were recorded and their photon burst histograms were plotted which demonstrated roughly the specific protein interaction ratio based on their population and statistical behavior. As a proof of concept, Src/STAT3 protein complex interaction ratios with and without EGF stimulation were obtained by MAPS within 1 h and the results were well matched with the one obtained by the conventional immunoprecipitation/Western blot (IP/WB).

authors

• Kameoka, Jun

published proceedings

• Analyst

author list (cited authors)

• Chou, C., Jing, N., Yamaguchi, H., Tsou, P., Lee, H., Chen, C., ... Hung, M.

citation count

• 9

publication date

• January 2010

publisher

• Royal Society of Chemistry (RSC)  Publisher

published in

• The Analyst  Journal

keywords

• Epidermal Growth Factor
• Fluorescent Dyes
• Hela Cells
• Humans
• Immunoglobulin G
• Microfluidic Analytical Techniques
• Protein Binding
• Quantum Dots
• STAT3 Transcription Factor
• Src-Family Kinases

Digital Object Identifier (DOI)

• 10.1039/c0an00229a

start page

• 2907

end page

• 2912

volume

• 135

issue

• 11

URL

• http://dx.doi.org/10.1039/c0an00229a