Development and characterization of novel T-cell receptor-like antibodies targeting human papillomavirus type 16...

Cervical cancer remains one of the most prevalent malignancies among women worldwide, with persistent infection by high-risk human papillomavirus (HPV), particularly types 16 and 18, as the principal etiological factor. The viral oncoproteins E6 and E7 are well-established drivers of cellular transformation and the maintenance of malignant phenotypes. This study aimed to generate T-cell receptor-like antibodies (TCR-like antibodies) capable of recognizing HPV16-derived E6 and E7 peptides presented by the major histocompatibility complex class I molecule HLA-A2402, with the goal of developing a diagnostic platform for early detection of HPV16-associated disease. Peptide-major histocompatibility complex (P-MHC) antigenic complexes were constructed and characterized using molecular docking, followed by experimental validation. A fully human domain antibody phage display library was screened against these complexes through iterative biopanning, supported by polyclonal and monoclonal enzyme-linked immunosorbent assays (ELISA). Structural evaluation and binding affinity analyses were performed using molecular docking, Western blotting, TCR-like antibody ELISA, and cell-based ELISA. Two novel TCR-like antibodies, designated D2\Ab and A12\Ab, were identified and demonstrated strong and specific binding to HPV16 E6 and E7 P-MHC complexes, confirming high affinity and antigen specificity. Functional validation across multiple assays demonstrated selective recognition of target complexes without reactivity toward irrelevant controls. In conclusion, this study demonstrates the feasibility of generating fully human TCR-like antibodies for HPV-associated malignancies and highlights their potential application in early detection and targeted diagnostic strategies for cervical cancer.