In vivo far-red luminescence imaging of a biomarker based on BRET from Cypridina bioluminescence to an organic dye.
We aimed to develop a far-red luminescence imaging technology for visualization of disease specific antigens on cell surfaces in a living body. First, we conjugated a far-red fluorescent indocyanine derivative to biotinylated Cypridina luciferase. This conjugate produced a bimodal spectrum that has long-wavelength bioluminescence emission in the far-red region as a result of bioluminescence resonance energy transfer. To generate a far-red luminescent probe with targeting and imaging capabilities of tumors, we then linked this conjugate to an anti-human Dlk-1 monoclonal antibody via the biotin-avidin interaction. This far-red luminescent probe enabled us to obtain high-resolution microscopic images of live, Dlk-1-expressing Huh-7 cells without an external light source, and to monitor the accumulation of this probe in tumor-bearing mice. Thus this far-red luminescent probe is a convenient analytical tool for the evaluations of monoclonal antibody localization in a living body.
Wu
Chun
C
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology, 1-8-31 Midorigaoka, Ikeda 563-8577, Japan.
Mino
Kazuhiro
K
Akimoto
Hidetoshi
H
Kawabata
Makiko
M
Nakamura
Koji
K
Ozaki
Michitaka
M
Ohmiya
Yoshihiro
Y
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015415:Biomarkers / Q000378:metabolism; D001710:Biotin; D002135:Calcium-Binding Proteins; D045744:Cell Line, Tumor; D003445:Crustacea / Q000201:enzymology; D004735:Energy Transfer; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D008156:Luciferases; D049449:Luminescence; D049408:Luminescent Agents; D008164:Luminescent Proteins / Q000235:genetics / Q000276:immunology; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009374:Neoplasms, Experimental / Q000378:metabolism; D013053:Spectrophotometry
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D002135:Calcium-Binding Proteins; C512941:DLK1 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D049408:Luminescent Agents; D008164:Luminescent Proteins; D008565:Membrane Proteins; C485195:luciferase, cypridina; D001710:Biotin; D008156:Luciferases