Figure 4.

General flow chart for the cloning, display and engineering of high-affinity Vβ neutralizing agents by yeast display. A Vβ clone that is specific for the SAg of interest is cloned into the yeast display vector (Figure 3) and used to generate libraries of mutants. The libraries are selected with fluorescently-labeled ligands, (e.g., conformation-specific anti-Vβ antibodies or the SAg of interest). Multiple rounds of selections are conducted to enrich the Vβ mutants with desired properties, which serve as templates for subsequent library design and screening to achieve desirable stability or affinity of Vβ with SAg.