Ligand Engineering via Yeast Surface Display and Adherent Cell Panning

Area of Science:

• Biotechnology
• Molecular Biology
• Drug Discovery

Background:

• High-throughput ligand discovery is crucial for targeted therapies, diagnostics, and basic science.
• Maintaining target antigen integrity, especially for membrane proteins, presents significant technical hurdles in selection processes.

Purpose of the Study:

• To present techniques for selecting target-binding ligands using yeast display on intact mammalian cells.
• To address challenges in target antigen presentation and non-specific binding during ligand selection.

Main Methods:

• Utilizing yeast-displayed ligand libraries panned on intact mammalian cells expressing the target antigen.
• Employing target-negative cells to deplete non-specific binders from the selected library.
• Implementing avidity reduction strategies to favor the selection of high-affinity ligands.

Main Results:

• Demonstrated the effectiveness of panning on intact cells for ligand selection against membrane targets.
• Successfully depleted non-specific binders using target-negative cell controls.
• Showcased avidity reduction as a method to enrich for high-affinity binders.

Conclusions:

• Panning yeast-displayed libraries on intact cells is a robust strategy for discovering ligands against challenging targets.
• The described techniques enhance the quality and specificity of selected ligands for therapeutic and diagnostic applications.