Antibody engineering to improve manufacturability

Area of Science:

• Biotechnology
• Protein Engineering
• Mammalian Cell Culture

Background:

• Antibody expression in Chinese Hamster Ovary (CHO) cells can vary significantly.
• A specific human monoclonal antibody, mAb B-c, exhibited poor expression and high aggregation levels, posing manufacturing challenges.

Purpose of the Study:

• To identify the source of manufacturability issues for mAb B-c.
• To engineer an alternative light chain (LC) to improve antibody expression and product quality.

Main Methods:

• Transient and stable expression in CHO-K1 cells were used to assess antibody production.
• Light chain shuffling was performed with the original heavy chain (HC) to identify novel LCs.
• Binding affinity, expression levels, and aggregate formation were evaluated.

Main Results:

• The light chain (LC) was identified as the primary cause of mAb B-c's manufacturability problems.
• Antibodies engineered with new LCs demonstrated high expression at a 1:1 or 2:1 LC:HC ratio, comparable to other antibodies.
• The new LCs significantly reduced high molecular weight aggregates and heavy chain (HC) dimer formation.

Conclusions:

• Light chain engineering is a viable strategy to overcome antibody manufacturability issues.
• Pairing heavy chains with alternative light chains can enhance antibody expression and product quality.
• This approach maintains or improves antibody binding affinity.