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 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.
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.|ARC Centre of Excellence for Nanoscale Biophotonics (CNBP), Macquarie University , Sydney, New South Wales 2109, Australia.
Nima
N
Sayyadi
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.|Minomic International Ltd. , Macquarie Park, Sydney, New South Wales 2113, Australia.
Irene
I
Justiniano
Department of Physics and Astronomy, Macquarie University , Sydney, New South Wales 2109, Australia.
Russell E
RE
Connally
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
Run
R
Zhang
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
Bingyang
B
Shi
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
Liisa
L
Kautto
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.|ARC Centre of Excellence for Nanoscale Biophotonics (CNBP), Macquarie University , Sydney, New South Wales 2109, Australia.
Arun V
AV
Everest-Dass
State Key Laboratory of Fine Chemicals, School of Chemistry, Dalian University of Technology , Dalian, Liaoning Province 116024, China.
Jingli
J
Yuan
Minomic International Ltd. , Macquarie Park, Sydney, New South Wales 2113, Australia.
Bradley J
BJ
Walsh
ARC Centre of Excellence for Nanoscale Biophotonics (CNBP), Macquarie University , Sydney, New South Wales 2109, Australia.|School of Mathematical and Physical Sciences, Faculty of Science, University of Technology , Sydney, New South Wales 2007, Australia.
Dayong
D
Jin
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
Robert D
RD
Willows
ARC Centre of Excellence for Nanoscale Biophotonics (CNBP), Macquarie University , Sydney, New South Wales 2109, Australia.|Department of Physics and Astronomy, Macquarie University , Sydney, New South Wales 2109, Australia.
James A
JA
Piper
Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.|ARC Centre of Excellence for Nanoscale Biophotonics (CNBP), Macquarie University , Sydney, New South Wales 2109, Australia.
Nicolle H
NH
Packer
D000911:Antibodies, Monoclonal / Q000276:immunology; D045744:Cell Line, Tumor; D056831:Coordination Complexes / Q000138:chemical synthesis / Q000494:pharmacology*; D005063:Europium / Q000737:chemistry*; D053673:Glypicans / Q000276:immunology; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D007158:Immunologic Techniques / Q000379:methods*; D008024:Ligands; D049449:Luminescence; D049408:Luminescent Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D008297:Male; D011471:Prostatic Neoplasms / Q000175:diagnosis*
D000911:Antibodies, Monoclonal; D056831:Coordination Complexes; C000621866:GPC1 protein, human; D053673:Glypicans; D018796:Immunoconjugates; D008024:Ligands; D049408:Luminescent Agents; D005063:Europium