:: Volume 4, Issue 3 (6-2017) ::
2017, 4(3): 34-38 Back to browse issues page
Isolation and purification of HLA-DR antigen from Daudi cell line by immunoaffinity chromatography
Zahra Khayyati , Fatemeh Yari
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran , f.yari@ibto.ir
Abstract:   (3854 Views)

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.

Keywords: HLA-DR, Affinity chromatography, ELISA
Full-Text [PDF 546 kb]   (1891 Downloads)    
Type of Study: Research | Subject: Immunology
Received: 2016/09/3 | Accepted: 2016/11/7 | Published: 2017/02/9

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Volume 4, Issue 3 (6-2017) Back to browse issues page