Near Infrared Fluorescence Imaging of EGFR Expression In Vivo Using IRDye800CW-nimotuzumab

Bernhard W, et al., 9:6213-6227, Oncotarget, 2018

Epidermal growth factor receptor (EGFR) is a cell surface glycoprotein and is often overexpressed in all aggressive cancers of epithelial origin. Nimotuzumab is an anti-EGFR antibody. Described herein is the development of a near infrared (NIR) fluorescent dye-labeled nimotuzumab immunoconjugate, IRDye800CW-nimotuzumab, and the evaluation of it as a useful NIR fluorescent imaging probe. Furthermore, in vitro and in vivo binding of IRDye-800CW-nimotuzumab to a variety of EGFR positive human cancer cell lines and ex vivo biodistribution was also investigated. Binding interactions between the antibodies and recombinant proteins were investigated using Bio-Layer Interferometry (BLI). A Pall ForteBio Octet RED384 instrument equipped with Anti-HumanFab-CH1 2nd Generation (FAB2G) Biosensor probes were used to perform assays based on BLI. Antibody immobilized FAB2G sensor tips were immersed in various concentrations of the target human EGFR proteins. BLI data obtained were fitted globally to a 1:1 binding model. The equilibrium dissociation constant (KD) was determined. Collectively, the results of this study suggest that nimotuzumab is a promising candidate for NIR fluorescent imaging and image-guided surgery.

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