Int. J. Mol. Sci. 2013, 14(9), 19146-19154; doi:10.3390/ijms140919146
Probing Antigen-Antibody Interaction Using Fluorescence Coupled Capillary Electrophoresis
1
School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou 213164, China
2
Department of Chemistry, the Chinese University of Hong Kong, Shatin, Hong Kong
â€
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Received: 23 August 2013 / Revised: 4 September 2013 / Accepted: 5 September 2013 / Published: 17 September 2013
(This article belongs to the Special Issue Molecular Bases of Cancer Research)
Abstract
In this report, the use of fluorescence detection coupled capillary electrophoresis (CE-FL) allowed us to fully characterize the antigen-antibody interaction. CE-FL allowed separation of unbound quantum dots (QDs) and ligand bound QDs and also revealed an ordered assembly of biomolecules on QDs. Further, we observed FRET from QDs donor to DyLight acceptor, which were covalently conjugated with human IgG and goat anti-human IgG, respectively. The immunocomplex was formed and the mutual affinity of the antigen and antibody brought QDs and DyLight close enough to allow FRET to occur. This novel CE-based technique can be easily extended to other FRET systems based on QDs and may have potential application in the detection of antibodies.Keywords:
QDs; FRET; immunocomplex; capillary electrophoresis
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Wang, J.; Qiu, L.; Wang, C.; Zhang, Y.; Li, J.; Xia, J.; Jiang, P. Probing Antigen-Antibody Interaction Using Fluorescence Coupled Capillary Electrophoresis. Int. J. Mol. Sci. 2013, 14, 19146-19154.
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