High-sensitivity quantum dot-based fluorescence resonance energy transfer bioanalysis by capillary electrophoresis.
Here a new method for high-sensitivity quantum dot (QD)-based fluorescence resonance energy transfer (FRET) bioanalysis was developed. In this method, capillary electrophoresis (CE) with fluorescence detection was applied. The FRET system consisted of water-soluble 532-nm emitting CdTe QDs donor and 632-nm emitting CdSe/ZnS QDs acceptor which were covalently conjugated with mouse IgG and goat anti-mouse IgG, respectively. The bio-affinity between antigen and antibody brought two kinds of QDs close enough to make the FRET happen between them. In the CE experiments, highly efficient separation of donor-acceptor immunocomplexes was obtained, and the process of FRET was monitored. Results showed that FRET efficiency obtained by CE (38.56-69.58%) improved substantially in comparison with that obtained by ensemble measurement (12.77-52.37%). The high efficient separation of donor-acceptor immunocomplexes and the possible conformation change of antigen and antibody, contributes to the lower analysis uncertainty (variance) and higher FRET efficiency obtained in CE and consequentially, this makes the analysis of FRET more sensitive. This novel CE-based technique can be easily extended to other FRET system based on QDs and may have potential application in the study of biomolecule conformation change.
Li
Yong-Qiang
YQ
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, Hubei, PR China.
Wang
Jian-Hao
JH
Zhang
Hai-Li
HL
Yang
Jie
J
Guan
Li-Yun
LY
Chen
Hong
H
Luo
Qing-Ming
QM
Zhao
Yuan-Di
YD
D015374:Biosensing Techniques / Q000295:instrumentation*; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation*; D007118:Immunoassay / Q000295:instrumentation*; D045663:Quantum Dots; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity