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1987
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ImmunologyPathologyPortfolio ManagementImmunotherapeuticsViral Structural ProteinCancer-associated VirusAutomobile InsuranceHuman Papillomavirus VaccinesRisk ManagementManagementInsurancePortfolio OptimizationVirologyPortfolio AllocationCell BiologyPortfolio InsuranceFinanceMolecular VirologyCervical CancerPathogenesisHpv16 L2Hpv CapsidMedicineViral OncologyViral ImmunityPrecancerous Lesions
Human papillomavirus infection is a leading cause of morbidity and mortality worldwide, causing benign warts and a range of malignancies including cervical cancer. The study aimed to produce recombinant HPV16 L1 and L2 proteins in 293‑F cells via transient co‑transfection. Recombinant proteins were expressed in 293‑F cells, purified by affinity chromatography, and formulated into L1/L2 VLPs that were used to immunize mice. The recombinant L1 and L2 proteins were successfully expressed, localized in nuclei and cytoplasm, formed VLPs visualized by TEM, and elicited higher anti‑L1 antibody titers in mice, indicating efficient transient expression and potential for vaccine development.
<h3>Abstract</h3> Human papillomavirus (HPV) infection is a leading cause of morbidity and mortality in women worldwide. The virus is associated with benign warts and a broad spectrum of malignancies, including cervical cancer, considered a disease of high clinical relevance, especially in developing countries. In this study we developed the production of recombinant proteins HPV16 L1 and HPV16 L2 in human cells in suspension (293-F), which were transiently co-transfected with the pUF3/L1h and pUF3/L2h vectors. Expressions of recombinant HPV16 L1 and L2 capsid proteins was detected by laser scanning confocal microscopy and flow cytometry. Both proteins were identified intracellularly in the nucleus and cytoplasm of cells. The presence of these heterologous proteins and VLPs formation were detected by transmission electron microscopy (TEM) through colloidal gold immunolabeling and negative staining. Cell extracts containing recombinant proteins were purified by affinity chromatography and immunization of Balb/c mice with the formulation HPV16 L1/L2 VLPs containing adjuvant was able to induce higher titer of anti-HPV16 L1, when compared to HPV16 L2 antibodies by indirect ELISA assay. These data indicate that transient expression in 293-F cells was efficiently established. The results are promising for obtain recombinant proteins of the HPV capsid for future studies involving human papillomavirus, as well as to contribute for the development of other vaccine strategies for prevention against HPV.