Development of positron emission tomography probe of 64Cu-labeled anti-C-kit 12A8 Fab to measure protooncogene C-kit expression

Introduction: C-kit is an important diagnostic and therapeutic target molecule for several malignancies, and c-kit-targeted drugs have been used clinically. Because abundant c-kit expression in tumors is a prerequisite for successful c-kit-targeted therapy, imaging of c-kit expression is expected to play a pivotal role in the therapeutic decision for each patient. We evaluated Cu-64-labeled Fab of anti-c-kit antibody 12A8 as a positron emission tomography (PET) imaging probe. Methods: In-111- or I-125-Labeled 12A8 Fab was evaluated in vitro by cell binding, competitive inhibition and cellular internalization assays, and in vivo by biodistribution in mice bearing c-kit-expressing and -non-expressing tumors. Next, Fab fragment was labeled with the positron emitter Cu-64 and evaluated by PET. Results: Radiolabeled 12A8 Fab showed specific binding to c-kit-expressing cells with high affinity and internalized into cells after binding to c-kit on cell surface. Although tumor accumulation of[In-111]Fab was lower than that of[In-111]IgG, the faster blood clearance of [In-111]Fab provided higher tumor-to-blood ratio at 6 h postinjection onwards. Blood clearance of Cu-64-labeled 12A8 Fab was slower than that of [In-111] Fab, but PET using [Cu-64, c Fab clearly visualized the tumor at 6 h postinjection onwards. Conclusion: The Cu-64-labeled 1 2A8 Fab could be used for c-kit-specific PET imaging and might help in selecting appropriate patients for c-kit-targeted treatments. (C) 2011 Elsevier Inc. All rights reserved.