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. Detectio...
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| Published in | Nature biotechnology Vol. 18; no. 7; pp. 764 - 767 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
New York, NY
Nature
01.07.2000
Nature Publishing Group |
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| Online Access | Get full text |
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| 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+/-0.2x10(-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. |
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| AbstractList | 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. 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 plus or minus 0.2 x 10 super(-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. |
| Audience | Academic |
| Author | Griffin, Guy D Cullum, Brian M Alarie, Jean-Pierre Vo-Dinh, Tuan |
| Author_xml | – sequence: 1 givenname: Tuan surname: Vo-Dinh fullname: Vo-Dinh, Tuan organization: Advanced Monitoring Development Group, Life Sciences Division, P.O. Box 2008, Oak Ridge National Laboratory – sequence: 2 givenname: Jean-Pierre surname: Alarie fullname: Alarie, Jean-Pierre organization: Advanced Monitoring Development Group, Life Sciences Division, P.O. Box 2008, Oak Ridge National Laboratory – sequence: 3 givenname: Brian M surname: Cullum fullname: Cullum, Brian M organization: Advanced Monitoring Development Group, Life Sciences Division, P.O. Box 2008, Oak Ridge National Laboratory – sequence: 4 givenname: Guy D surname: Griffin fullname: Griffin, Guy D organization: Advanced Monitoring Development Group, Life Sciences Division, P.O. Box 2008, Oak Ridge National Laboratory |
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| Keywords | Carcinogen Antibody Metabolite Biosensor DNA In situ Molecular probe Intracellular Lesion Measurement method Nanotechnology |
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| References_xml | – year: 1991 ident: b1 article-title: Fiber optic chemical sensors and biosensors contributor: fullname: Wolfbeis, O.S. – volume: 64 start-page: 2985 year: 1992 end-page: 2990 ident: b8 publication-title: Anal. Chem. contributor: fullname: Kopelman, R. – volume: 3 start-page: 85 year: 1993 end-page: 92 ident: b7 publication-title: Immunomethods contributor: fullname: Alarie, J.P. – year: 1989 ident: b13 article-title: Chemical analysis of polycyclic aromatic compounds contributor: fullname: Vo-Dinh, T. – volume: 8 start-page: 45 year: 1996 end-page: 52 ident: b5 publication-title: Polycyclic Aromatic Compounds contributor: fullname: Vo-Dinh, T. – volume: 70 start-page: 2646 year: 1998 end-page: 2650 ident: b12 publication-title: Anal. Chem. contributor: fullname: Vo-Dinh, T. – volume: 37 start-page: 532 year: 1991 end-page: 535 ident: b6 publication-title: Clin. Chem. contributor: fullname: Santella, R.M. – volume: 251 start-page: 1468 year: 1991 end-page: 1470 ident: b11 publication-title: Science contributor: fullname: Kostelak, R.L. – volume: 258 start-page: 778 year: 1992 end-page: 781 ident: b9 publication-title: Science contributor: fullname: Kopelman, R. – volume: 41 start-page: 735 year: 1987 end-page: 738 ident: b4 publication-title: Appl. Spectrosc. contributor: fullname: Vo-Dinh, T. – volume: 236 start-page: 237 year: 1990 end-page: 244 ident: b14 publication-title: Anal. Chim. Acta contributor: fullname: Vo-Dinh, T. – year: 1992 ident: b3 article-title: Immunochemical assays and biosensor technology contributor: fullname: Rechnitz, G.A. – volume: 5 start-page: 373 year: 1984 end-page: 377 ident: b15 publication-title: Carcinogenesis contributor: fullname: Weinstein, I.B. – year: 1996 ident: b2 contributor: fullname: Harmer, A. – volume: 71 start-page: 2071 year: 1999 end-page: 2075 ident: b10 publication-title: Anal. Chem. contributor: fullname: Kopelman, R. – volume: 251 start-page: 1468 year: 1991 ident: BFnbt0700_764_CR11 publication-title: Science doi: 10.1126/science.251.5000.1468 contributor: fullname: E Betzig – volume-title: Fiber optic chemical sensors and biosensors year: 1991 ident: BFnbt0700_764_CR1 contributor: fullname: OS Wolfbeis – volume: 236 start-page: 237 year: 1990 ident: BFnbt0700_764_CR14 publication-title: Anal. Chim. Acta doi: 10.1016/S0003-2670(00)83317-6 contributor: fullname: JP Alarie – volume: 3 start-page: 85 year: 1993 ident: BFnbt0700_764_CR7 publication-title: Immunomethods doi: 10.1006/immu.1993.1042 contributor: fullname: T Vo-Dinh – ident: BFnbt0700_764_CR2 – volume: 258 start-page: 778 year: 1992 ident: BFnbt0700_764_CR9 publication-title: Science doi: 10.1126/science.1439785 contributor: fullname: T Tan – volume-title: Chemical analysis of polycyclic aromatic compounds year: 1989 ident: BFnbt0700_764_CR13 contributor: fullname: T Vo-Dinh – volume: 5 start-page: 373 year: 1984 ident: BFnbt0700_764_CR15 publication-title: Carcinogenesis doi: 10.1093/carcin/5.3.373 contributor: fullname: RM Santella – volume-title: Immunochemical assays and biosensor technology year: 1992 ident: BFnbt0700_764_CR3 contributor: fullname: RM Nakamura – volume: 70 start-page: 2646 year: 1998 ident: BFnbt0700_764_CR12 publication-title: Anal. Chem. doi: 10.1021/ac971304f contributor: fullname: V Deckert – volume: 64 start-page: 2985 year: 1992 ident: BFnbt0700_764_CR8 publication-title: Anal. Chem. doi: 10.1021/ac00047a019 contributor: fullname: W Tan – volume: 71 start-page: 2071 year: 1999 ident: BFnbt0700_764_CR10 publication-title: Anal. Chem. doi: 10.1021/ac9901081 contributor: fullname: SLR Barker – volume: 8 start-page: 45 year: 1996 ident: BFnbt0700_764_CR5 publication-title: Polycyclic Aromatic Compounds doi: 10.1080/10406639608048333 contributor: fullname: JP Alarie – volume: 37 start-page: 532 year: 1991 ident: BFnbt0700_764_CR6 publication-title: Clin. Chem. doi: 10.1093/clinchem/37.4.532 contributor: fullname: T Vo-Dinh – volume: 41 start-page: 735 year: 1987 ident: BFnbt0700_764_CR4 publication-title: Appl. Spectrosc. doi: 10.1366/0003702874448076 contributor: fullname: T Vo-Dinh |
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| SubjectTerms | 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 |
| Title | Antibody-based nanoprobe for measurement of a fluorescent analyte in a single cell |
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