Metastatic castration-resistant prostate cancer (mCRPC) presents a significant challenge of global health. This study aims to establish non-invasive liquid biopsy assays for Neurotensin (NTS) to detect mCRPC and its in disease progression.

Monoclonal antibodies (mAbs) for NTS using hybridoma and phage display techniques were generated, followed by characterization through ELISA and Western blotting. We isolated medium derived from mCRPC cell lines (PC3 and C4-2B) to detect NTS expression. In vitro proliferation assays and in vivo xenograft models were employed to evaluate the biological relevance of NTS expression in mCRPC.

Our assays identified high-affinity mAbs targeting NTS, with elevated expression detected in mCRPC cells, mCRPC xenograft, and clinical samples. We used a full-length NTS peptide (13 amino acids) to screen for anti-NTS monoclonal antibodies from a phage display library. We identified six antibodies for evaluation of their binding affinity using ELISA.

This study underscores the clinical utility of NTS as biomarkers for theranostics in mCRPC. The developed liquid biopsy assays offer a non-invasive tool to monitor disease progression and guide personalized treatment strategies.