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Antibody-based nanoprobe for measurement of a fluorescent analyte in a single cell

Abstract

We report here the application of an antibody-based nanoprobe for in situ measurements of a single cell. The nanoprobe employs antibody-based receptors targeted to a fluorescent analyte, benzopyrene tetrol (BPT), a metabolite of the carcinogen benzo[a]pyrene (BaP) and of the BaP–DNA adduct. Detection of BPT is of great biomedical interest, since this species can serve as a biomarker for monitoring DNA damage due to BaP exposure and for possible precancer diagnosis. The measurements were performed on the rat liver epithelial clone 9 cell line, which was used as the model cell system. Before making measurements, the cells were treated with BPT. Nanoprobes were inserted into individual cells, incubated 5 min to allow antigen–antibody binding, and then removed for fluorescence detection. We determined a concentration of 9.6 ± 0.2 × 10−11 M for BPT in the individual cells investigated. The results demonstrate the possibility of in situ measurements inside a single cell using an antibody-based nanoprobe.

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Figure 1: Development of nanoprobes.
Figure 2: Scanning electron microscope image of a nanofiber.
Figure 3: Optical measurement system.
Figure 4: Photograph of an antibody-based nanoprobe used to measure presence of BPT in a single cell.
Figure 5: Calibration measurements and single-cell sensing using the nanoprobes after various exposures.

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Acknowledgements

This research is sponsored by the LDRD Project (Advanced Nanosensors) and by the Office of Biological and Environmental Research, US Department of Energy, under contract DE-AC05-960R22464 with Lockheed Martin Energy Research Corp.

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Correspondence to Tuan Vo-Dinh.

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Vo-Dinh, T., Alarie, JP., Cullum, B. et al. Antibody-based nanoprobe for measurement of a fluorescent analyte in a single cell. Nat Biotechnol 18, 764–767 (2000). https://doi.org/10.1038/77337

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