Development and Use of IgM/J‐Chain Fusion Proteins for Characterization of Immunoglobulin Superfamily Ligand‐Receptor Interactions. Issue 1 (3rd February 2014)

• Record Type: Journal Article
• Authors: Ammann, Johannes U.; Trowsdale, John
• Editors: Coligan, John E.; Dunn, Ben M.; Speicher, David W.; Wingfield, Paul T.
• Is Part Of: Current protocols in protein science. Volume 75:Issue 1(2014)
• Page Start: 19.24.1
• Page End: 19.24.11
• Publication Date: 2014-02-03
• Subjects: immunoglobulin -- ligand‐receptor interaction -- fusion proteins -- IgM; Proteins -- Laboratory manuals; Proteins; Clinical Laboratory Techniques; Genetic Techniques; Immunologic Techniques; Proteins; Laboratory manuals; 572.6028
• DOI: 10.1002/0471140864.ps1924s75
• Languages: English
• ISSNs: 1934-3655

Abstract

Abstract: Discovery of binding partners for immunoglobulin molecules expressed by cells of the immune system is an important topic of current research. However, many ligand‐receptor interactions are of low affinity, and hence detection is refractory to most established protocols. We evaluated fusion proteins based on human IgM as high avidity probes to screen for ligand‐receptor binding. We describe methods for cloning, expression, and quantification of IgM fusion proteins with J‐chain. Furthermore, we outline protocols to assess binding of IgM fusion proteins to cells and to plate‐bound proteins. Compared to standard IgG‐fusion proteins, IgM + J chain increased binding of a test interaction, PD‐L1 to PD‐1, up to 1000‐fold. Curr. Protoc. Protein Sci . 75:19.24.1‐19.24.11. © 2014 by John Wiley & Sons, Inc.