Isolation of Antibodies to Heparan Sulfate on Glypicans by Phage Display. (9th August 2018)

• Record Type: Journal Article
• Authors: Kim, Heungnam; Ho, Mitchell
• Editors: Coligan, John E.; Dunn, Ben M.; Speicher, David W.; Wingfield, Paul T.
• Is Part Of: Current protocols in protein science. Volume 94(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-08-09
• Subjects: glypican -- heparan sulfate proteoglycan -- phage display -- single‐chain variable fragment or scFv; Proteins -- Laboratory manuals; Proteins; Clinical Laboratory Techniques; Genetic Techniques; Immunologic Techniques; Proteins; Laboratory manuals; 572.6028
• DOI: 10.1002/cpps.66
• Languages: English
• ISSNs: 1934-3655

Abstract

Abstract: Heparan sulfate (HS) plays an important role in development and disease. It interacts with many growth factors, chemokines, and other ligands known to be important for cell growth, motility, and differentiation. However, isolating an antibody to HS in mice, rabbits, or humans is difficult due to the poor immunogenicity of HS. Phage display is a major antibody engineering technology that allows the selection of antibodies for poorly immunogenic or highly conserved antigens. This protocol contains detailed procedures for HS antigen preparation and isolation of a phage displayed human single‐chain Fv (HS20) that binds HS on glypican‐3 (GPC3), and analysis of the selected phage antibody. It is conceivable that the procedures described in this protocol may be applicable to the isolation of antibodies for a variety of HS molecules. © 2018 by John Wiley & Sons, Inc.