Targeted dual selection to optimize transposon stable pool generation of multispecifics

Area of Science:

• Biotechnology
• Molecular Biology
• Protein Engineering

Background:

• Next-generation biologics, including bispecific and trispecific antibodies, leverage multispecific targeting for enhanced efficacy.
• The complexity of these molecules presents significant challenges in expression and purification, often resulting in low yields and mispairing.
• Stable cell pool expression is crucial for advancing these complex molecules from research to development.

Purpose of the Study:

• To develop and implement an optimized method for generating stable cell pools for complex multispecific antibodies.
• To address the limitations of low yields and mispairing in the purification of bispecific and trispecific antibodies.
• To enhance the production and quality of designer molecules for biopharmaceutical research.

Main Methods:

• Implemented targeted dual selection (TDS), a novel approach combining selective pressure with transposon-guided gene integration.
• Utilized analytical data from high-throughput transient productions to predict optimal vector configurations.
• Generated and evaluated optimized TDS stable pools for bispecific and trispecific antibody production.

Main Results:

• Achieved up to a four-fold increase in protein of interest yields for Y-shaped bispecific and cross-over dual variable trispecific molecules.
• Maintained product quality during the initial capture purification step.
• Demonstrated improved molecule quality at the initial capture purification stage.

Conclusions:

• Optimized stable cell pools using TDS can significantly improve yields and quality for complex multispecific antibody production.
• This strategy simplifies purification processes and increases production yields, facilitating timely project progression.
• TDS offers a valuable approach for advancing the research and development of next-generation biologics.