© Springer International Publishing Switzerland 2015. Preclinical data in fluorescence-guided surgery have provided strong evidence that real-time navigation for cancer localization is highly feasible. However, many of these cancer-specific probes are likely to be expensive and time-consuming to translate to the clinic. To facilitate the translation of cancer-specific localization of fluorescence for surgical navigation, therapeutic antibodies are being coupled with fluorescent molecules to provide robust contrast between disease and normal tissue. In this chapter, specific components of the antibody-based approach are detailed to provide an overview of the requirements for successful translation into human use.