Monoclonal antibody as ligand for purification of rabies virus protein from the brain of infected dogs and mice

Immunoaffinity chromatography using monoclonal antibodies (mAbs) as ligands has been used to purify rabies virus (RV) individual proteins. In this method, mAbs against RV were firstly purified, coupled to CnBr-agarose resin and used for purification RV individual proteins. Brain tissue homogenates derived from infected and uninfected dogs and mice were mixed with mAbs-CnBr agarose resin and washed extensivelly phosphate buffered salin (PBS). Following elution and neutralization, purified proteins were detected by enzyme-linked immunosirbent assay (ELISA) and Western blotting assay. Of the three mAbs (BB5, AE11 and AF6) as ligands, mAb AE11-CnBr agarose resin yielded highest protein levels as compared to those of mAb BB5-CnBr agarose and mAb AF6-CnBr agarose resins. In Western Blotting assay, the purified protein appeared to be 65 Kda (glycoprotein) and 38 kDa proteins. In ELISA test, the purified proteins reacted with both mAbs and policlonal antibodies (pAbs). Kata kunci: CnBr-agarose, rabies virus, protein, purificaion, chromaography