title: Activatable fluorescent cys-diabody conjugated with indocyanine green derivative: consideration of fluorescent catabolite kinetics on molecular imaging.
abstract: Antibody fragments including diabodies have more desirable pharmacokinetic characteristics than whole antibodies. An activatable optical imaging probe based on a cys-diabody targeting prostate-specific membrane antigen conjugated with the near-infrared fluorophore, indocyanine green (ICG), was designed such that it can only be activated when bound to the tumor, leading to high signal-to-background ratios. We employed short polyethylene glycol (PEG) linkers between the ICG and the reactive functional group (Sulfo-OSu group), resulting in covalent conjugation of ICG to the cys-diabody, which led to lower dissociation of ICG from cys-diabody early after injection, reducing hepatic uptake. However, unexpectedly, high and long-term fluorescence was observed in the kidneys, liver, and blood pool more than 1 h after injection of the cys-diabody PEG-ICG conjugate. A biodistribution study using I125-labeled cys-diabody-ICG showed immediate uptake in the kidneys followed by a rapid decrease, while gastric activity increased due to released radioiodine during rapid cys-diabody-ICG catabolism in the kidneys. To avoid this catabolic pathway, it would be preferable to use antibody fragments large enough not to be filtered through glomerulus or to conjugate the fragments with fluorescent dyes that are readily excreted into urine when cleaved from the cys-diabody to achieve high tumor-specific detection.
authors.affiliation: National Institutes of Health, National Cancer Institute, Center for Cancer Research, Molecular Imaging Program, Bethesda, Maryland 20892, USA.
authors.forename: Kohei
authors.initials: K
authors.lastname: Sano
authors.forename: Takahito
authors.initials: T
authors.lastname: Nakajima
authors.forename: Towhid
authors.initials: T
authors.lastname: Ali
authors.forename: Derek W
authors.initials: DW
authors.lastname: Bartlett
authors.forename: Anna M
authors.initials: AM
authors.lastname: Wu
authors.forename: Insook
authors.initials: I
authors.lastname: Kim
authors.forename: Chang H
authors.initials: CH
authors.lastname: Paik
authors.forename: Peter L
authors.initials: PL
authors.lastname: Choyke
authors.forename: Hisataka
authors.initials: H
authors.lastname: Kobayashi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000378:metabolism; D000954:Antigens, Surface / Q000276:immunology / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000493:pharmacokinetics; D043425:Glutamate Carboxypeptidase II / Q000276:immunology / Q000378:metabolism; D007208:Indocyanine Green / Q000737:chemistry* / Q000493:pharmacokinetics; D007700:Kinetics; D051379:Mice; D008819:Mice, Nude; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D011092:Polyethylene Glycols; D014018:Tissue Distribution
chemical_list: D000906:Antibodies; D000954:Antigens, Surface; D005456:Fluorescent Dyes; D011092:Polyethylene Glycols; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II; D007208:Indocyanine Green