1. Similar to phage display, yeast display provides a direct connection between genotype and phenotype. A plasmid containing the gene of interest is contained within yeast cells and the encoded antibody is expressed on the cell surface.

2.The display level of each yeast cell is variable, with each cell displaying 104 to 105 copies of the VHH.

3. Screening of antibody variation of surface expression and avidity can be quantified using fluorescence-activated cell sorting (FACS), which measures both the strength of antigen binding and amount of antibody expression on the yeast cell surface using separate tags on the antibody and antigen.

4. FACS binding assays provide a much more quantitative way of assessing high binding affinity and selectivity for the antigens and offer the ability to accurately control selection parameters (binding population percentage, signal normalization, and desired binding affinities) by flow cytometry.

5.There are usually 1-2 rounds of enrichment of target antigen-binding clones.

6. The antibody displayed by a single yeast colony is evaluated for specificity and reproducibility. If the antibody has the required functionality, the antibody gene is sequenced as part of antibody validation.