In CHO cells, the expression levels of heavy chain (HC) and light chain (LC) in monoclonal antibodies (mAb), and the subunits in bispecific antibodies, have a significant impact on productivity and product quality. Studies have shown that the modulation of HC and LC expression levels directly influences antibody productivity and quality. We would like to share our experiences with dosage compensation in the production of biologics.

HCPs in adeno-associated virus (AAV) products can be effectively enriched by ProtoeMiner beads and the detergent Pluronic F-68 can be simultaneously removed without loss of low-abundance HCPs.

Up to 34-fold increase in the enrichment of HCPs can be achieved by using ProteoMiner beads comparing to direct digestion.

After applying ProtoeMiner beads on AAV products, HCPs at a level as low as 0.1 ng/mL can be detected.

• Strategies To Balancing Chain Ratio and Assembly Issues To Maximise Formation of Fully Formed Multispecifics;
• Solutions for preventing light chain swapping;
• Analytical techniques to screen and ensure proper chain pairing;
• Case studies on successful mitigation of chain mismatching issues;
• Can transposons give a better ratio?
• Vector design strategies? Promoters? - Explore innovative strategies for optimizing expression vectors, with a focus on chain ratio modulation;
• Keep product impurities to a minimum;
• Strategies to navigate longer timelines and additional assays required for multispecifics;
• How to reduce aggregation of product within the cell? Bioreactor conditions? Engineering?

Targeted integration (TI) approaches, which involve integration of a transgene into a specific locus in the genome, are increasingly utilized for CHO cell line development (CLD) in recent years. But none of these CLD approaches are suitable for expression of toxic or difficult-to-express molecules, or can help in determining the underlying causes for a poor expression molecule. Here we introduce how a regulated target integration (RTI) system can help determine the underlying causes of low protein expression in an antibody (mAb-A). In addition, we will discuss how using a RTI system can help boost specific productivity and protein expression without negatively impacting cell growth.

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