Construction and screening of antigen targeted immune yeast surface display antibody libraries.
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.
Miller
Keith D
KD
Pacific Northwest National Laboratory, Richland, Washington, USA.
Pefaur
Noah B
NB
Baird
Cheryl L
CL
D000465:Algorithms; D000818:Animals; D000917:Antibody Formation / Q000235:genetics / Q000502:physiology; D000937:Antigen-Antibody Reactions / Q000276:immunology; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000378:metabolism*; D008280:Magnetics; D019151:Peptide Library; D015003:Yeasts / Q000235:genetics / Q000276:immunology*
D000954:Antigens, Surface; D007128:Immunoglobulin Fragments; D019151:Peptide Library