Proteolytic processing of recombinant anti-PD-1 monoclonal antibody through ectopic expression of the PACE enzyme in CHO-DG44 cells.

• Grant number: 23/00684-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2023
• End date: December 31, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Mari Cleide Sogayar
• Grantee: Matheus Costa Lourenço
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 16/05311-2 - Regenerative medicine aiming at therapy for chronic degenerative diseases (cancer and diabetes), AP.TEM

Abstract

In view of the benefits of immunotherapy for the treatment of cancer, compared to conventional treatments, in recent years, the search for new immunotherapies has gained increasing importance. New production strategies aim to improve the performance and effectiveness of these immunotherapeutic drugs.In this project, a new expression strategy for a monoclonal antibody that acts on the PD-1/PD-L1 system will be studied, using a bicistronic construction to express the light and heavy chains of the antibody in the same mRNA. Its ectopic expression, performed in CHO cells of the DG44 lineage, will have post-translational processing assisted by the endoprotease Furin/PACE. Thus, the aim is to generate a recombinant CHO-DG44 cell line that stably expresses the PACE enzyme so that these cells are then transformed with the pNU1 vector containing the antibody coding sequences.