UNIT 19.24 Development and Use of IgM/J-Chain Fusion Proteins for Characterization of Immunoglobulin Superfamily Ligand-Receptor Interactions
Published Online: 3 FEB 2014
Copyright © 2013 John Wiley & Sons, Inc. All rights reserved.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Ammann, J. U. and Trowsdale, J. 2014. Development and Use of IgM/J-Chain Fusion Proteins for Characterization of Immunoglobulin Superfamily Ligand-Receptor Interactions. Current Protocols in Protein Science. 75:19.24:19.24.1–19.24.11.
- Published Online: 3 FEB 2014
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.
- ligand-receptor interaction;
- fusion proteins;