isolation and purification of hla-dr antigen from daudi cell line by immunoaffinity chromatography

Introduction: the major histocompatibility complex (mhc) is a group of cell surface proteins that are essential for recognizing foreign molecules in human and other mammals. the physiologic function of mhc molecules is the presentation of peptides to t cells. in this study, we evaluated the purification of a class ii mhc molecule (hla-dr) from a human burkitt′s lymphoma cell line; daudi. materials and methods: we described a simple procedure for purifying human hla molecules from the cells lysate. as a representative model, hla-dr was purified from daudi cell line. the cell membrane was solubilized by a buffer contained np-40 detergent. subsequently, the isolation of the membrane antigen was carried out by affinity chromatography method using mouse anti-human hla-dr monoclonal antibody. the size and the specificity of the purified antigen were determined by bradford and elisa methods, respectively. results: the purified hla antigen was obtained in approximately 20-30 micrograms in each run of chromatography. additionally, elisa method demonstrated the hla-dr specificity of the purified protein. conclusion: the results indicated that affinity purification of hla-dr antigen by means of specific monoclonal antibody is a simple and fast procedure for obtaining the purified antigen.