Isolation and purification of HLA-DR antigen from Daudi cell line by immunoaffinity chromatography
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Zahra Khayyati , Fatemeh Yari  |
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran , f.yari@ibto.ir |
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Abstract: (4463 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. |
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Keywords: HLA-DR, Affinity chromatography, ELISA |
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Full-Text [PDF 546 kb]
(2075 Downloads)
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Type of Study: Research |
Subject:
Immunology Received: 2016/09/3 | Accepted: 2016/11/7 | Published: 2017/02/9
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