A novel intracellular nanobody against HPV16 E6 oncoprotein

Cervical cancer occurs as a result of the persistent infection of high-risk human papillomavirus (HPV). HPV16 oncoproteins E6 and E7 exert different and concerted pro-tumor actions in cell transformation and malignance maintenance in various m echanisms. Nanobody expressed as “intracellular antibodies” (intrabodies) can target intracellular antigens to hamper their function efficaciously and specifically. In this work, phage-display approach was employed to select the high affinity HPV16 E6-specific nanobody, nanobody Nb9 against HPV16 E6 was selected. Nb9 has high affinity (Kaff =6.3 × 108 M−) and can specifically bind endogenous HPV16 E6 protein in HPV16 positive CaSki and SiHa cells. In Nb9 overexpressed SiHa and CaSki cells, nucleus localization of HPV16 E6 was inhibited, p53 inactivation was prevented and increased apoptosis was observed. Moreover, tumor growth was inhibited in mouse xenograft model. Taken together, our results suggested that nanobody Nb9 could be a useful inhibitor for HPV16 E6 function and particularly appropriate for the treatment of HPV-associated disease. •Phage display nanobody library was constructed and HPV16 E6-specific nanobody Nb9 was selected by panning.•Binding specificity and affinity to HPV16 E6 was determined with purified E.coli expressed Nb9 protein.•Nb9 intrabody reduced proliferation of HPV16-positive tumor cells in vitro.•In vivo, Nb9 intrabody expression inhibited tumor growth in HPV16 mouse models.