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

• Published in: Nature biotechnology Vol. 18; no. 7; pp. 764 - 767
• Main Authors: Vo-Dinh, Tuan, Alarie, Jean-Pierre, Cullum, Brian M, Griffin, Guy D
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.07.2000; Nature Publishing Group
• Subjects: Animals; Antibodies; Antibodies - immunology; Benzo(a)pyrene - metabolism; benzopyrene tetrol; Biological and medical sciences; Biomarkers; Biosensors; Biotechnology; Carcinogens; Cell Line; Chemical mutagenesis; Cloning; Deoxyribonucleic acid; DNA; DNA Adducts - metabolism; DNA damage; Epithelium - metabolism; Fluorescence; Fluorescent Antibody Technique - instrumentation; Fundamental and applied biological sciences. Psychology; In situ measurement; Liver - metabolism; Medical sciences; Metabolites; Methods. Procedures. Technologies; Microscopy, Electron, Scanning; Nanotechnology; Pyrene; Pyrenes - metabolism; Rats; Silver; Spectrometry, Fluorescence - instrumentation; Spectrometry, Fluorescence - methods; Toxicology; Various methods and equipments; Carcinogen; Antibody; Metabolite; Biosensor; DNA; In situ; Molecular probe; Intracellular; Lesion; Measurement method; Nanotechnology
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/77337

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+/-0.2x10(-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.