Antibody-based nanoprobe for measurement of a fluorescent analyte in a single cell.
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.
Vo-Dinh
T
T
Advanced Monitoring Development Group, Life Sciences Division, P.O. Box 2008, Oak Ridge National Laboratory, Oak Ridge TN 37831-6101, USA.
Alarie
J P
JP
Cullum
B M
BM
Griffin
G D
GD
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001564:Benzo(a)pyrene / Q000378:metabolism; D015415:Biomarkers; D002460:Cell Line; D018736:DNA Adducts / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000295:instrumentation*; D008099:Liver / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D011721:Pyrenes / Q000378:metabolism; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods*
D000906:Antibodies; D015415:Biomarkers; D018736:DNA Adducts; D011721:Pyrenes; C406414:benzo(a)pyrene tetrol; D001564:Benzo(a)pyrene