New Biotin Derivative-DOTA Conjugate
J ournal of Medicinal Chemistry, 2003, Vol. 46, No. 14 3173
activity aspecifically bound to the Centricon, an 90Y-labeled
r-BHD aliquot was centrifuged without adding Av. All runs
were performed in duplicate.
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Sta bility Stu d ies. Aliquots from the labeling mixture were
diluted with saline or human serum/saline 1:1 solution alter-
natively. After 2, 6, 12, 24, 36, and 48 h of incubation at 37
°C, two aliquots (10 µL) of both the mixtures were drawn: one
was added to Av molar excess and the other was added to
saline. The samples were centrifuged in Centricon YM-10 and
subsequently washed with 3 × 200 µL saline. The presence of
90Y-labeled r-BHD/Av complex, free 90Y ion, and biotinidase-
cleaved 90Y-DOTA, potentially generated during incubation,
was determined by the activity counts of the top and bottom
parts of the Centricon with and without Av excess (see above).
Ack n ow led gm en t. This work was supported in part
by the Italian Association for Cancer Research (AIRC).
The content of this work is covered by Italian Patent
Application RM2001A000079.
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Ap p en d ix
Abbreviations: Av, avidin; DOTA, 1,4,7,10-tetraaza-
cyclododecane-1,4,7,10-tetraacetic acid; EDC, 1-(3-dim-
ethylaminopropyl)-3-ethylcarbodiimide hydrochloride;
HATU, 1-[bis(dimethylamino)methylene]-1H-1,2,3-tria-
zolo(4,5-b)pyridinium 1-hexafluorophosphate 3-oxide,
O-(7-benzotriazol-1-yl)-1,1,3,3,-tetramethyluronium hexa-
fluorophosphate; NMM, N-methylmorpholine; Sav,
streptavidin; sulfo-NHS, N-hydroxysulfosuccinimide so-
dium salt, monosodium 1-hydroxy-2,5-dioxo-3-pyrro-
lidinesulfonate.
Su p p or tin g In for m a tion Ava ila ble: 1H NMR data, more
detail of the radiolabeling procedure, and biological studies
as well as elemental analyses data are available free of charge
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