Enhanced Production of HCV E1E2 Subunit Vaccine Candidates via Protein-Protein Interaction Identification in Glyco-engineered CHO cells

Hepatitis C Virus (HCV) is a bloodborne virus that affects 70 million people globally with infections that can often go unnoticed, thus motivating the development of an HCV vaccine. While prior work has developed secreted E1E2 (sE1E2) protein vaccines, efforts to express it recombinantly have resulted in very low titers. To address this challenge, here we employed a multi-omics approach to identify protein interactors that enhance the secretion of sE1E2. By detecting Protein-Protein Interactions (PPIs) using Biotinylation by Antibody Recognition (BAR) and integrating the data with RNA-Seq, we identified proteins within the secretory pathway that interact with sE1E2 and validated their impact by overexpressing the interacting proteins. Among these, CUL4A and YWHAH significantly enhanced sE1E2 secretion in geCHO cells. The integration of omics techniques and genetic engineering in this study provides valuable insights into improving protein secretion in CHO cells, paving the way for the development of more affordable and accessible biotherapeutics. Overall design: RNA-seq profiling of glyco-engineered CHO cells expressing the subunit HCV vaccine, E1E2