Sensitive Time-Gated Immunoluminescence Detection of Prostate Cancer Cells Using a TEGylated Europium Ligand

We describe the application of a synthetically developed tetradentate β-diketonate-europium chelate with high quantum yield (39%), for sensitive immunodetection of prostate cancer cells (DU145). MIL38 antibody, a mouse monoclonal antibody against Glypican 1, conjugated directly to the chelate via ly...

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Published in:Analytical chemistry (Washington) 2016-10, Vol.88 (19), p.9564-9571
Main Authors: Sayyadi, Nima, Justiniano, Irene, Connally, Russell E., Zhang, Run, Shi, Bingyang, Kautto, Liisa, Everest-Dass, Arun V., Yuan, Jingli, Walsh, Bradley J., Jin, Dayong, Willows, Robert D., Piper, James A., Packer, Nicolle H.
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Language:English
Subjects:
Antibodies
Cancer
Cells
Chelates
Conjugates
Europium
Luminescence
Marking
Polyclonal antibodies
Prostate
Prostate cancer
Rodents
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cited_by cdi_FETCH-LOGICAL-a409t-8b3d8e92084a7fa3bb31629cb921d1b675baeb21941cf212491b6d7199d417233
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container_issue 19
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container_title Analytical chemistry (Washington)
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creator Sayyadi, Nima
Justiniano, Irene
Connally, Russell E.
Zhang, Run
Shi, Bingyang
Kautto, Liisa
Everest-Dass, Arun V.
Yuan, Jingli
Walsh, Bradley J.
Jin, Dayong
Willows, Robert D.
Piper, James A.
Packer, Nicolle H.
description We describe the application of a synthetically developed tetradentate β-diketonate-europium chelate with high quantum yield (39%), for sensitive immunodetection of prostate cancer cells (DU145). MIL38 antibody, a mouse monoclonal antibody against Glypican 1, conjugated directly to the chelate via lysine residues, resulted in soluble (hydrophilic) and stable immunoconjugates. Indirect labeling of the antibody by a europium chelated secondary polyclonal antibody and a streptavidin/biotin pair was also performed. All of these bright luminescent conjugates were used to stain DU145 cells, a prostate cancer cell line, using time gated luminescence microscopy for imaging, and their performances were compared to conventional FITC labeling. For all prepared conjugates, the europium chelate in conjunction with a gated autosynchronous luminescence detector (GALD) completely suppressed the cellular autofluorescence background to allow capture of vivid, high contrast images of immune-stained cancer cells.
doi_str_mv 10.1021/acs.analchem.6b02191
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source American Chemical Society:Jisc Collections:American Chemical Society Read & Publish Agreement 2022-2024 (Reading list)
subjects Antibodies
Cancer
Cells
Chelates
Conjugates
Europium
Luminescence
Marking
Polyclonal antibodies
Prostate
Prostate cancer
Rodents
title Sensitive Time-Gated Immunoluminescence Detection of Prostate Cancer Cells Using a TEGylated Europium Ligand
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