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Study on molecular interactions between proteins on live cell membranes using quantum dot-based fluorescence resonance energy transfer

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Abstract

Mouse anti-human CD71 monoclonal antibody (anti-CD71) was conjugated with red quantum dots (QDs; 5.3 nm, emission wavelength λ em = 614 nm) and used to label HeLa cells successfully. Then green QD-labeled goat anti-mouse immunoglobulin G (IgG; the size of the green QDs was 2.2 nm; λ em = 544 nm) was added to bind the red-QD-conjugated anti-CD71 on the cell surface by immunoreactions. Such interaction between anti-CD71 and IgG lasted 4 min and was observed from the fluorescence spectra: the fluorescence intensity of the “red” peak at 614 nm increased by 32%; meanwhile that of the “green” one at 544 nm decreased by 55%. The ratio of the fluorescence intensities (I 544 nm/I 614 nm) decreased from 0.5 to 0.2. The fluorescence spectra as well as cell imaging showed that fluorescence resonance energy transfer took place between these two kinds of QDs on the HeLa cells through interactions between the primary antibody and the secondary antibody.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (grants no. 30670553, 30370387) and the Program for Distinguish Young Scientist of Hubei Province (2006ABB020).

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Authors and Affiliations

  1. Key Laboratory of Biomedical Photonics of Ministry of Education - Wuhan National Laboratory for Optoelectronics - Hubei Bioinformatics and Molecular Imaging Key Laboratory, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, HuBei, 430074, China

    Tian-Cai Liu, Hai-Li Zhang, Jian-Hao Wang, Hai-Qiao Wang, Zhi-Hong Zhang, Xiao-Feng Hua, Yuan-Cheng Cao, Qing-Ming Luo & Yuan-Di Zhao

  2. Department of Physics, National University of Singapore, 2 Science Drive 3, Singapore, 117542, Singapore

    Tian-Cai Liu

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  1. Tian-Cai Liu
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  2. Hai-Li Zhang
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Correspondence to Yuan-Di Zhao.

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Liu, TC., Zhang, HL., Wang, JH. et al. Study on molecular interactions between proteins on live cell membranes using quantum dot-based fluorescence resonance energy transfer. Anal Bioanal Chem 391, 2819–2824 (2008). https://doi.org/10.1007/s00216-008-2189-3

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  • DOI: https://doi.org/10.1007/s00216-008-2189-3

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