Trastuzumab or TDM1 were obtained from the MSK Hospital Pharmacy. The pHrodo-TDM1 was obtained by conjugating the free lysine residues of TDM1 with the amine-reactive pH-sensitive pHrodo iFL Red STP ester dye (ThermoFisher Scientific, P36014) according to the manufacturer’s instructions.
To prepare [89Zr]Zr-DFO-antibody, TDM1 or Trastuzumab were first conjugated with the bifunctional chelate p-isothiocyanatobenzyl-desferrioxamine (DFO-Bz-NCS; Macrocyclics, Inc) and then labeled with zirconium-89 (89Zr)59 (link). Radiochemical purity (RCP) was determined by instant thin-layer chromatography. The radiolabeled conjugates used for in vitro and in vivo studies had a RCP of 99%, radiochemical yields ranging from 92 to 97%, specific activities in the range of 21.98–24.73 Mbq/nmol, and immunoreactivities above 90%.
To prepare [89Zr]Zr-DFO-antibody, TDM1 or Trastuzumab were first conjugated with the bifunctional chelate p-isothiocyanatobenzyl-desferrioxamine (DFO-Bz-NCS; Macrocyclics, Inc) and then labeled with zirconium-89 (89Zr)59 (link). Radiochemical purity (RCP) was determined by instant thin-layer chromatography. The radiolabeled conjugates used for in vitro and in vivo studies had a RCP of 99%, radiochemical yields ranging from 92 to 97%, specific activities in the range of 21.98–24.73 Mbq/nmol, and immunoreactivities above 90%.
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