Sensitive Time-Gated Immunoluminescence Detection of Prostate Cancer Cells Using a TEGylated Europium Ligand
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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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Abstract
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.
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Analytical Chemistry
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88
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19
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Analytical chemistry
Other chemical sciences
Biochemistry and cell biology not elsewhere classified
Medical biochemistry and metabolomics
Chemical engineering