Expression of Bispecific Antibodies Using Cellular Display

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Biology

Background:

• Selecting high-producing cell lines is often resource-intensive and time-consuming.
• Assessing individual cell expression potential within a mixed population can streamline screening.

Purpose of the Study:

• To describe protocols for a cellular display technology to simplify cell line selection.
• To detail construct design and methods for tuning cellular display strength.
• To provide enrichment methods for flow cytometric sorting based on expression or product quality.

Main Methods:

• Utilizing alternate splicing to generate two mRNA constructs at tunable ratios.
• Designing constructs where one mRNA codes for the secreted product and the other directs the product to the cellular membrane via a transmembrane domain.
• Employing flow cytometry for sorting cell populations based on cellular display quantity or product quality attributes like bispecific antibody heteromerization.

Main Results:

• Protocols for cellular display technology are presented.
• Methods for tuning the strength of cellular display are detailed.
• Enrichment methods enable sorting based on expression levels and product quality.

Conclusions:

• Cellular display technology offers a simplified approach to screening and selecting high-producing cell lines.
• The described methods allow for efficient tuning and sorting based on desired expression and product quality attributes.