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Iranian Journal of Basic Medical Sciences، جلد ۲۰، شماره ۳، صفحات ۳۲۷-۳۳۳
عنوان فارسی
چکیده فارسی مقاله
کلیدواژه‌های فارسی مقاله
عنوان انگلیسی Selection of single-chain variable fragments specific for Mycobacterium tuberculosis ESAT-6 antigen using ribosome display
چکیده انگلیسی مقاله Objective(s): Tuberculosis (TB) is still one of the problematic infectious diseases in developing countries, especially in Iran. In the present study, we applied ribosome display technique to select single chain variable fragments (scFvs) specific for the 6-kDa early secretory antigenic target (ESAT-6) antigen of Mycobacterium tuberculosis from a mouse scFv library. Materials and Methods: The gene encoding ESAT-6 was cloned into pET22b(+) plasmid and expressed in Escherichia coli BL21 (DE3). The purified recombinant ESAT-6 protein was injected into female BALB/c mice for immunization, and then m-RNA was extracted from the spleen of immunized mice. The anti-ESAT-6 VH/k chain library was assembled by joining of VH and k into the VH/k chain with a 72-bp DNA linker by SOE (splicing by overlap extension) PCR. The scFv library was panned against ESAT-6 using a single round of ribosome display via a rabbit reticulocyte lysate system. Results: ELISA assay showed that one of the selected scFvs had higher affinity against the recombinant ESAT-6 protein. The affinity of the candidate scFv was ̴ 3.74×108 M-1. Conclusion: It could be proposed that the isolated scFv in this study may be useful for the diagnosis of TB.
کلیدواژه‌های انگلیسی مقاله
نویسندگان مقاله شهرزاد آهنگرزاده | shahrzad ahangarzadeh
department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

مژگان بنده پور | mojgan bandehpour
cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, iran|department of biotechnology, school of advanced technologies in medicine, shahid beheshti university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

بهرام کاظمی | bahram kazemi
department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran, iran|cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, iran|department of biotechnology, school of advanced technologies in medicine, shahid beheshti university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_8363.html
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/87/article-87-325103.pdf
کد مقاله (doi)
زبان مقاله منتشر شده en
موضوعات مقاله منتشر شده
نوع مقاله منتشر شده Original Article
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