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" Determination of Optimized Condition for Production of Camel Antibody (Nanobody) Against TIM-3 Using Phage Display Technology "
\ Vida Homayouni
; Abbas Rezaei, MazdakGanjaliKhaniHakemi
; Mahdi Behdani
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Isfahan University of Medical Sciences
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| Document Type
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Latin Dissertation
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| Language of Document
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English
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| Record Number
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109245
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| Doc. No
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T17320
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| Call number
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QW601,H763d,2016
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| Main Entry
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Homayouni, Vida
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| Title & Author
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Determination of Optimized Condition for Production of Camel Antibody (Nanobody) Against TIM-3 Using Phage Display Technology\ Vida Homayouni
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| College
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Schools, Medical
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| Date
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, 2016
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| Degree
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Ph.D
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| field of study
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Immunology
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| Page No
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67p.: ill, diag, tab
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| Note
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This is a research dissertation with project ID: 392603
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ویدا همایونی
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| Abstract
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As T-cell immunoglobulin and mucin domain 3 (TIM-3) is an immune regulatorymolecule; blocking or stimulating of itcould alter the pattern of immune responses towards desired condition.Based on the unique features of Nanobodies, we aimed toconstruct an anti-TIM-3 Nanobody as an appropriate tool for manipulating immune responses for future therapeutic purposes.A camel was immunized with TIM-3 antigen andthen, VHH phagmid library was constructed from its B cell’s genome using nested PCR. Library selection against TIM-3antigen was performed in three rounds of panning. Using phage-ELISA, the most reactive clones 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 flow cytometric analysis was performed to analyze the binding and biologic activitiesof theTIM-3 specific Nanobody with HL-60 and HEK cell lines expressed TIM-3.Specific 15kD band representing for Nanobody was observed in gel and confirmed with Western blotting. The Nanobody showed significant specific immune-reactivity against TIM-3 with a relatively high binding affinity. The Nanobodysignificantly suppressed the proliferation of TIM-3 expressing HL-60 cell line.For the first time, we successfully prepared afunctional anti-humanTIM-3 specific Nanobodywith a high affinityand an anti-proliferative activity on an AML cell line in vitro.
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| Descriptor
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Immunoglobulin Domains
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Hepatitis A Virus Cellular Receptor 1
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Camelus
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| Added Entry
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Rezaei, Abbas, Thesis advisor
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GanjaliKhaniHakemi, Mazdak, Thesis advisor
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Behdani , Mehdi, Consulting advisor
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| Translated Title Supplied by Cataloguer
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تعیین شرایط بهینه تولید نانوبادی اختصاصی مولکول تنظیم کننده ایمنی TIM-3 با استفاده از تکنیک نمایش فاژی
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