Akademik Veri Yönetim Sistemi
International Journal of Peptide Research and Therapeutics, cilt.31, sa.6, 2025 (SCI-Expanded, Scopus)
Purpose: The ongoing COVID-19 pandemic has highlighted the urgent need for efficient and scalable recombinant protein vaccine production platforms. In this study, we developed and optimized a UCOE-based lentiviral vector system for the expression of target SARS-CoV-2 antigens in CHO cells. Method: Next-generation UCOE constructs and control plasmid vectors were obtained, and relevant antigen-coding sequences were PCR-amplified to generate stock DNA for cloning. Successful amplification and integrity of PCR products were verified by agarose gel electrophoresis, and proper insertion into plasmids was confirmed through restriction enzyme digestion analyses. HEK293T cells were co-transfected with antigen-bearing UCOE plasmids and lentiviral packaging vectors to produce lentiviral vectors (LVs), which were harvested at 48 and 72 h post-transfection. Viral titers quantified by flow cytometry confirmed efficient vector production. CHO cells transduced with optimized LV doses stably expressed recombinant RBD and SC2N antigens. Result: HPLC analysis revealed homogeneous peak profiles for both proteins, and measured LOD and LOQ values confirmed their purity and integrity, indicating high-quality antigen production. Q-TOF peptide mapping further validated the correct sequences of both RBD and SC2N proteins, confirming their structural integrity and indicating minimal undesired modifications. Expression kinetics showed RBD levels peaking at 72 h in culture medium, while SC2N peaked at 48 h in cells. Conclusion: Importantly, UCOE technology enhanced expression efficiency and reduced the likelihood of undesired post-translational modifications. These results demonstrate that the developed UCOE-based lentiviral vector system enables efficient and high-quality recombinant COVID-19 antigen production in CHO cells, supporting its potential as a scalable platform for vaccine development.