Abstract
Recombinant scfv antibodies specific for CYP1A1 and CYP1B1 P450 enzymes were combined with targeted imaging mass spectrometry to simultaneously detect the P450 enzymes present in archived, paraffin-embedded, human breast cancer tissue sections. By using CYP1A1 and CYP1B1 specific scfv, each coupled to a unique reporter molecule (i.e., a mass tag) it was possible to simultaneously detect multiple antigens within a single tissue sample with high sensitivity and specificity using mass spectrometry. The capability of imaging multiple antigens at the same time is a significant advance that overcomes technical barriers encountered when using present day approaches to develop assays that can simultaneously detect more than a single antigen in the same tissue sample.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / chemistry
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Antibodies, Monoclonal / immunology
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Aryl Hydrocarbon Hydroxylases / chemistry
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Aryl Hydrocarbon Hydroxylases / immunology
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Avidin / chemistry
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Avidin / immunology
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Biotin / chemistry
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Biotin / immunology
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Breast / chemistry
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Breast Neoplasms / chemistry
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Cytochrome P-450 CYP1A1 / chemistry
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Cytochrome P-450 CYP1A1 / immunology
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Cytochrome P-450 CYP1B1
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Enzyme-Linked Immunosorbent Assay
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Female
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Histocytochemistry / methods
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Humans
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Mass Spectrometry / methods*
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Mice
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Molecular Imaging / methods*
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Rats
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Sensitivity and Specificity
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Single-Chain Antibodies / chemistry*
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Single-Chain Antibodies / metabolism
Substances
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Antibodies, Monoclonal
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Recombinant Proteins
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Single-Chain Antibodies
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Avidin
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Biotin
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Aryl Hydrocarbon Hydroxylases
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CYP1A1 protein, human
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CYP1B1 protein, human
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Cyp1b1 protein, mouse
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Cyp1b1 protein, rat
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Cytochrome P-450 CYP1A1
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Cytochrome P-450 CYP1B1