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Iranian Journal of Basic Medical Sciences، جلد ۱۹، شماره ۱۱، صفحات ۱۲۰۱-۱۲۰۸
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عنوان انگلیسی Preparation and characterization of a novel nanobody against T-cell immunoglobulin and mucin-3 (TIM-3)
چکیده انگلیسی مقاله Objective(s): As T-cell immunoglobulin and mucin domain 3 (TIM-3) is an immune regulatory molecule; its blocking or stimulating could alter the pattern of immune response towards a desired condition. Based on the unique features of nanobodies, we aimed to construct an anti-TIM-3 nanobody as an appropriate tool for manipulating immune responses for future therapeutic purposes. Materials and Methods:We immunized a camel with TIM-3 antigen and then, synthesized a VHH phagemid library from its B cell’s transcriptome using nested PCR. Library selection against TIM-3antigen was performed in three rounds of panning. Using phage-ELISA, the most reactive colonies were selected for sub-cloning in soluble protein expression vectors. The Nanobody was purified and confirmed with a nickel-nitrilotriacetic acid (Ni-NTA) column, SDS-PAGE and Western blotting. A flowcytometric analysis was performed to analyze the binding and biologic activities of theTIM-3 specific nanobody with TIM-3 expressing HL-60 and HEK cell lines. Results:Specific 15kD band representing for nanobody was observed on the gel and confirmed with Western blotting. The nanobody showed significant specific immune-reactivity against TIM-3 with a relatively high binding affinity. The nanobody significantly suppressed the proliferation of TIM-3 expressing HL-60 cell line. Conclusion: Finally, we successfully prepared a functional anti-humanTIM-3 specific nanobody with a high affinity and an anti-proliferative activity on an AML cell line in vitro.
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نویسندگان مقاله ویدا همایونی | vida homayouni
immunology department, faculty of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

مزدک گنجعلی خانی حاکمی | mazdak ganjalikhani hakemi
immunology department, faculty of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

عباس رضایی | abbas rezaei
immunology department, faculty of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

حسین خان احمد | hossein khanahmad
genetic department, faculty of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

مهدی بهدانی | mahdi behdani
biotechnology research center, biotechnology department, venom amp;amp; bio-therapeutics molecules lab, pasteur institute of iran, tehran, iran

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

فاطمه کاظمی lomedasht | fatemeh kazemi lomedasht
biotechnology research center, biotechnology department, venom amp;amp; bio-therapeutics molecules lab, pasteur institute of iran, tehran, iran

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_7820.html
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