Construction and Screening of Antigen Targeted Immune Yeast Surface Display Antibody Libraries

Abstract These protocols describe a yeast surface display–based process for the rapid selection of antibodies from immunized mice, eliminating the need for creating and screening hybridoma fusions. A yeast surface display library of single‐chain antibody fragments (scFvs) is created from antigen‐binding B cells from the splenocytes of immunized mice. The antigen targeted library is then screened for antigen specific scFv by magnetic‐activated cell sorting (MACS) and fluorescence‐activated cell sorting (FACS). Library construction and screening can be accomplished in as little as 2 weeks, resulting in a panel of scFvs specific for the target antigen. Curr. Protocol. Cytom . 45:4.7.1‐4.7.30. © 2008 by John Wiley & Sons, Inc.