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 magneticactivated 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.
Revised: September 23, 2009 |
Published: August 2, 2009
Citation
Miller K.D., N.B. Pefaur, and C.L. Baird. 2009.Construction and Screening of Antigen Targeted Immune Yeast Surface Display Antibody Libraries. In Current Protocols in Cytometry. New York:Wiley.PNNL-SA-63652.