Unit

UNIT 4.7 Construction and Screening of Antigen Targeted Immune Yeast Surface Display Antibody Libraries

  1. Keith D. Miller1,
  2. Noah B. Pefaur2,
  3. Cheryl L. Baird1

Published Online: 1 JUL 2008

DOI: 10.1002/0471142956.cy0407s45

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Miller, K. D., Pefaur, N. B. and Baird, C. L. 2008. Construction and Screening of Antigen Targeted Immune Yeast Surface Display Antibody Libraries. Current Protocols in Cytometry. 45:4.7:4.7.1–4.7.30.

Author Information

  1. 1

    Pacific Northwest National Laboratory, Richland, Washington

  2. 2

    Department of Protein Biochemistry, ZymoGenetics, Seattle, Washington

Publication History

  1. Published Online: 1 JUL 2008
  2. Published Print: JUL 2008

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.

Keywords:

  • surface display;
  • yeast;
  • antibody;
  • immune library;
  • scFv