Non-small cell lung cancers (NSCLCs) constitute the majority of lung cancer cases and are associated with high mortality rates. Conventional therapies—including chemotherapy, radiotherapy, and targeted biological treatments—often provide limited clinical benefit and are frequently accompanied by substantial adverse effects. Therefore, the development of more effective and selective therapeutic strategies is urgently needed. In this study, we present a novel CD166-targeting monoclonal antibody (mAb) conjugated with dual payloads (DualADC) as a promising therapeutic agent for NSCLC. We first screened a new receptor CD166 in majority of NSCLC patients and developed an anti-CD166 monoclinal antibody (mAb). This mAb showed high NSCLC specificity and targeting efficiency in both in vitro and in vivo. Then we developed a novel conjugation platform and constructed a DualADC by linking CD166 mAb with a cytotoxic payload (Dxd) and an immune-stimulatory TLR7/8 agonist (modified R848). The scalable biomanufacturing process, including fed-batch production in a stirred-tank bioreactor and purification was established. Flow cytometry analysis confirmed its high binding affinity, specificity and cytotoxicity of our CD166-targeting DualADC across three NSCLC subtypes. In vivo anti-NSCLC studies validated the tumor burden reduction, metastasis inhibition and efficacy to treat heterogenous patient-derived xenograft. Further mechanism study demonstrated synergistic mechanism of actions (chemo- and immune- therapeutic functions) and minimal systemic toxicity (high specificity). In conclusion, our study introduces a new target and ADC for NSCLC treatment.
