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Cancer Biother Radiopharm. 2007 Oct;22(5):654-64.

Preparation, characterization, and biological evaluation of a streptavidin-chimeric t84.66 conjugate for antibody pretargeting.

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Department of Veterinary Medicine and Surgery, College of Veterinary Medicine and Surgery, University of Missouri-Columbia, Columbia, MO 65211, USA.


In order to develop a new system for the antibody pretargeting of carcinoembryonic antigen (CEA)-positive cancers, a streptavidin (SA) conjugate of the monoclonal antibody (mAb) chimeric, T84.66, was synthesized and characterized. Antibody disulfide bonds were reduced with 1,4-dithiothreitol, and the resulting thiols were reacted with succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate-derivatized streptavidin (SMCC-SA). The desired SA-cT84.66 conjugate was purified by iminobiotin affinity chromatography and size-exclusion high-performance liquid chromatography (HPLC). The molecular weight of the SA-cT84.66 conjugate (210 kDa) and immounoreactivity (100%) were confirmed by size-exclusion HPLC, and the conjugate bound three equivalents of (111)In-DOTA-biotin. SA-cT84.66-pretargeted (111)In-DOTA-biotin was evaluated in nude mice bearing LS174T human colon carcinoma xenografts. Tumor uptake of (111)In-DOTA-biotin peaked at 3.32% injected dose per gram after 15 minutes. Clearance from blood and normal organs was extremely rapid, and tumor-to-blood ratios were >20:1 after 24 hours. The specific tumor targeting and rapid whole-body clearance of SA-cT84.66-pretargeted (111)In-DOTA-biotin indicated that this system is promising for the imaging and therapy of CEA-positive cancers.

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