title: Simple and Efficient Production and Purification of Mouse Myelin Oligodendrocyte Glycoprotein for Experimental Autoimmune Encephalomyelitis Studies.
abstract: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS), thought to occur as a result of autoimmune responses targeting myelin. Experimental autoimmune encephalomyelitis (EAE) is the most common animal model of CNS autoimmune disease, and is typically induced via immunization with short peptides representing immunodominant CD4+T cell epitopes of myelin proteins. However, B cells recognize unprocessed protein directly, and immunization with short peptide does not activate B cells that recognize the native protein. As recent clinical trials of B cell-depleting therapies in MS have suggested a role for B cells in driving disease in humans, there is an urgent need for animal models that incorporate B cell-recognition of autoantigen. To this end, we have generated a new fusion protein containing the extracellular domain of the mouse version of myelin oligodendrocyte glycoprotein (MOG) as well as N-terminal fusions of a His-tag for purification purposes and the thioredoxin protein to improve solubility (MOGtag). A tobacco etch virus (TEV) protease cleavage site was incorporated to allow the removal of all tag sequences, leaving only the pure MOG1-125 extracellular domain. Here, we describe a simple protocol using only standard laboratory equipment to produce large quantities of pure MOGtag or MOG1-125. This protocol consistently generates over 200 mg of MOGtag protein. Immunization with either MOGtag or MOG1-125 generates an autoimmune response that includes pathogenic B cells that recognize the native mouse MOG.
authors.affiliation: Department of Microbiology and Immunology, Western University.
authors.forename: Rajiv W
authors.initials: RW
authors.lastname: Jain
authors.affiliation: Department of Microbiology and Immunology, Western University.
authors.forename: Amy K
authors.initials: AK
authors.lastname: Dang
authors.affiliation: Department of Microbiology and Immunology, Western University; skerfoot@uwo.ca.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Kerfoot
mesh_terms: D000818:Animals; D004681:Encephalomyelitis, Autoimmune, Experimental; D006801:Humans; D051379:Mice; D009103:Multiple Sclerosis; D018992:Myelin-Associated Glycoprotein; D063308:Myelin-Oligodendrocyte Glycoprotein; D013601:T-Lymphocytes
chemical_list: C571327:Mog protein, mouse; D018992:Myelin-Associated Glycoprotein; D063308:Myelin-Oligodendrocyte Glycoprotein