{"title":"Isolation and purification of HLA-DR antigen from Daudi cell line by immunoaffinity chromatography","authors":"Zahra Khayyati, F. Yari","doi":"10.29252/JBRMS.4.3.34","DOIUrl":null,"url":null,"abstract":"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.","PeriodicalId":15047,"journal":{"name":"Journal of Basic Research in Medical Sciences","volume":"4 1","pages":"34-38"},"PeriodicalIF":0.0000,"publicationDate":"2017-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Basic Research in Medical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.29252/JBRMS.4.3.34","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
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.