Circumventing Therapeutic Resistance and the Emergence of Disseminated Breast Cancer Cells Through Non-Invasive Optical Imaging.

摘要

Herein we explore a series of optically distinct near infrared emissive polymersomes (NIREPs; biodegradable polymer vesicles that manifest extraordinarily high irradiances and are ideally suited for in vivo optical imaging), each conjugated to different antibodies for the non-invasive molecular imaging of all breast cancer sites within a patient, including micrometastases. In Year 2, we have taken significant steps to optimize NIREP fabrication protocols to ensure antibodies can be reproducibly conjugated to the surface of NIREPs while preserving native antigen-recognition properties. We have developed a fluoronitrobenzoic acid (FNB) conjugation strategy, which enables us to covalently attach trastuzumab (anti-HER2 IgG) to the surface of pre-fabricated NIREPs under mild, aqueous conditions. In model experiments, we have demonstrated that covalent modification of trastuzumab does not affect HER2 recognition properties. Furthermore, we have developed a sensitive Western blot protocol for the detection of trastuzumab attached to NIREPs to allow accurate quantification of NIREP surface functionalization density. With our new protocols, we are well placed in Year 3 to generate a cocktail of antibody-functionalized NIREPs for in vitro and in vivo evaluation.