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 ce...

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Bibliographic Details
Published inCurrent protocols in cytometry Vol. Chapter 4; p. Unit4.7
Main Authors Miller, Keith D, Pefaur, Noah B, Baird, Cheryl L
Format Journal Article
LanguageEnglish
Published United States 01.07.2008
Subjects
Algorithms
Animals
Antibody Formation - genetics
Antibody Formation - physiology
Antigen-Antibody Reactions - immunology
Antigens, Surface - genetics
Antigens, Surface - immunology
Antigens, Surface - metabolism
B-Lymphocytes - immunology
Cell Separation - methods
Cloning, Molecular - methods
Humans
Immunoglobulin Fragments - genetics
Immunoglobulin Fragments - metabolism
Magnetics
Peptide Library
Yeasts - genetics
Yeasts - immunology
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Summary: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.
ISSN:1934-9300
DOI:10.1002/0471142956.cy0407s45