Isolation and purification of HLA-DR antigen from Daudi cell line by immunoaffinity chromatography

نویسندگان

  • Fatemeh Yari Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
  • Zahra Khayyati Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
چکیده مقاله:

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.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

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. materi...

متن کامل

Isolation of chitin deacetylase from Mucor rouxii by immunoaffinity chromatography

The purification of chitin deacetylase from Mucor rouxii to homogeneity employing conventional methods has already been described. However, a lengthy protocol is required resulting in a low yield and specific activity for the enzyme . A 169-fold one-step purification of chitin deacetylase by immunoaffinity chromatography is reported, resulting in a homogeneous enzyme preparation. The enzyme pur...

متن کامل

Purification of nucleotide-requiring enzymes by immunoaffinity chromatography.

Monospecific (affinity-purified) anti-(yeast glucose-6-phosphate dehydrogenase) IgG inhibits three different NADPH-requiring enzymes, chicken liver dihydrofolate reductase, pigeon liver fatty acid synthetase and chicken liver malic enzyme. The inhibition of all three enzymes was approx. 50% in a 2h incubation with 100 micrograms of IgG. Similarly, with several different NADH-requiring enzymes, ...

متن کامل

Purification of TEM-1 beta-lactamase by immunoaffinity chromatography.

A monoclonal antibody prepared against TEM-1 beta-lactamase was found to compete with penicillins and cephalosporins for binding to the enzyme. The purified antibody preparation was linked to Sepharose 4B and used for immunoaffinity-chromatography purification of TEM-1 beta-lactamase. Elution with either benzylpenicillin or cloxacillin yielded a highly purified, concentrated and active enzyme p...

متن کامل

One-step purification of nisin A by immunoaffinity chromatography.

The lantibiotic nisin A was purified to homogeneity by a single-step immunoaffinity chromatography method. An immunoadsorption matrix was developed by direct binding of anti-nisin A monoclonal antibodies to N-hydroxysuccinimide-activated Sepharose. The purification procedure was rapid and reproducible and rendered much higher final yields of nisin than any other described method.

متن کامل

Isolation of Coccidioides immitis F antigen by immunoaffinity chromatography with monospecific antiserum.

Detection of antibody to Coccidioides immitis F antigen is of proved value in the diagnosis of coccidioidomycosis. This antibody is demonstrable by use of an immunodiffusion assay with reference coccidioidin antigen and antiserum to C. immitis. Using a combination of lectin affinity and immunoaffinity chromatography, we isolated the F antigen from coccidioidin and prepared monospecific antibody...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

ذخیره در منابع من قبلا به منابع من ذحیره شده

{@ msg_add @}


عنوان ژورنال

دوره 4  شماره 3

صفحات  34- 38

تاریخ انتشار 2017-06

با دنبال کردن یک ژورنال هنگامی که شماره جدید این ژورنال منتشر می شود به شما از طریق ایمیل اطلاع داده می شود.

کلمات کلیدی

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023