Improving Pertuzumab production by gene optimization and proper signal peptide selection

Area of Science:

• Biotechnology
• Molecular Biology
• Protein Engineering

Background:

• Optimizing protein expression in Chinese Hamster Ovary (CHO) cells is crucial for therapeutic antibody production.
• Signal peptide selection and codon optimization are key factors influencing protein secretion efficiency.

Purpose of the Study:

• To enhance Pertuzumab production by optimizing heavy and light chain genes and selecting appropriate signal peptides.
• To evaluate the impact of sequence optimization and signal peptides on Pertuzumab secretion and antigen binding.

Main Methods:

• Transient transfection of CHO-K1 cells with optimized and non-optimized antibody gene constructs.
• Employing five different signal peptides to assess secretion enhancement.
• Analyzing Pertuzumab production rates and binding affinity to ErbB2 antigen.

Main Results:

• Whole heavy and light chain sequence optimization resulted in a 3.8-fold increase in Pertuzumab production.
• Human albumin signal peptide showed a two-fold increase in monoclonal antibody production, though not statistically significant.
• Signal peptide selection did not affect Pertuzumab's binding to the ErbB2 antigen.

Conclusions:

• Combined whole antibody sequence optimization and human albumin signal peptide effectively improve Pertuzumab production rates.
• This optimized sequence facilitates the development of stable Pertuzumab-producing CHO-K1 cell lines.
• The findings support the efficient production of Pertuzumab as a biosimilar drug.