Development and Use of IgM/J‐Chain Fusion Proteins for Characterization of Immunoglobulin Superfamily Ligand‐Receptor Interactions

Autor:	Johannes U. Ammann, John Trowsdale
Rok vydání:	2014
Předmět:	Immunoassay Cloning Recombinant Fusion Proteins Biology Ligand (biochemistry) Biochemistry Molecular biology Fusion protein J chain HEK293 Cells Immune system Immunoglobulin M Structural Biology biology.protein Humans Immunoglobulin superfamily Cloning Molecular Antibody Receptor
Zdroj:	Current Protocols in Protein Science
ISSN:	1934-3663 1934-3655
DOI:	10.1002/0471140864.ps1924s75
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::6404d993bd1ca1598cfe3d9119177587 https://doi.org/10.1002/0471140864.ps1924s75 Zobrazit plný text záznamu Plný text