Analysis of Mannan (Polymannose)-Peptide Conjugate by Competitive ELISA

Multiple sclerosis (MS) is an autoimmune disease whereby the myelin of the central nervous system (CNS) is destroyed, leading to paralysis and serious health problems [1].The main proteins of the membrane sheath of myelin are: i) myelin basic protein (MBP); ii) myelin oligodendrocyte glycoprotein (MOG), iii) myelin-associated glycoprotein (MAG) and iv) proteolipid protein (PLP) [2,3].Immunodominant epitopes of myelin proteins are involved in pathogenesis of MS.Thus, the 35-55 sequence of MOG protein is an autoantigen of MS and induces in vivo Experimental Autoimmune Encephalomyelitis (EAE-animal model of MS) [3].Moreover, the conjugate of this epitope with mannan (polysaccharide) was found to inhibit the EAE symptoms and could be a new and promising approach for MS treatment [4,5].In the present study, a competitive ELISA was developed using the MOG36-55 peptide for the coating of polystyrene microtiter plates, polyclonal antibodies produced in rabbits after immunization with this peptide, and the mannan-MOG conjugates as competitors.The first approach was to test the stability of mannan-MOG conjugates during storage conditions and their potency after modifications at the peptide sequence that may occur spontaneously.The developed methodology is possible to be used in further analysis of mannan -peptide conjugate in combination with in vivo experiments