We performed synthesis and testing of Fab’-based NIR fluorescent probes with the goal of achieving sufficiently high tumor/background (muscle) ratios at the earlier time points after intravenous injection in comparison to the full-size IgG or larger fragments. Conjugation of NIRDye 800CW to Fab’ resulted in partially quenched imaging probes. Fluorescence intensity imaging of orthotopic xenografts in both EGFR-attenuated and wild-type triple negative breast cancer tumors using Fab’-800CW allowed earlier detection of EGFR expression in vivo.
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