Optimizing Expression of Multi-chain and Bispecific Antibodies: Strategies and Considerations

Expression Platforms:
Traditional expression platforms optimized for monoclonal antibodies (mAbs) need to be adapted for the growing prevalence of multi-chain asymmetric antibodies and bispecifics (bsAbs).

Vector Optimization:
The design of high-efficiency expression vectors, including bicistronic constructs with internal ribosome entry sites (IRESs), can significantly improve the production of recombinant antibodies in CHO cells.

Metabolic Engineering:
Metabolic engineering approaches, such as two-stage selection strategies, can help develop stable cell lines with high-quality mAb production by controlling metabolic activity and ensuring long-term stability.

Cell Line Selection:
CHO cells are the most commonly used host for therapeutic mAb production due to their high expression levels and adaptability to large-scale production.

Transient vs. Stable Expression:
Both transient and stable expression methodologies have their advantages, with transient expression offering rapidity and economy, while stable expression provides long-term stability and high productivity.

Cell Culture Optimization:
Manipulating cell growth pathways, such as apoptosis and metabolic pathways, and using additives like growth factors and histone deacetylase inhibitors can enhance antibody expression in cell lines.

Bacterial Antibody Display (BAD):
BAD technology can be used to identify framework variants that improve antibody expression and thermostability, offering a valuable tool for optimizing antibody production.
By leveraging these strategies, researchers can overcome the challenges associated with the expression of multi-chain and bispecific antibodies, leading to more efficient and cost-effective production of these therapeutic agents.