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center : Isfahan University of Medical Sciences
Document Type : Latin Dissertation
Language of Document : English
Record Number : 109245
Doc. No : T17320
Call number : QW601,H763d,2016
Main Entry : Homayouni, Vida
Title & Author : Determination of Optimized Condition for Production of Camel Antibody (Nanobody) Against TIM-3 Using Phage Display Technology\ Vida Homayouni
College : Schools, Medical
Date : , 2016
Degree : Ph.D
field of study : Immunology
Page No : 67p.: ill, diag, tab
Note : This is a research dissertation with project ID: 392603
: ویدا همایونی
Abstract : 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.
Descriptor : Immunoglobulin Domains
: Hepatitis A Virus Cellular Receptor 1
: Camelus
Added Entry : Rezaei, Abbas, Thesis advisor
: GanjaliKhaniHakemi, Mazdak, Thesis advisor
: Behdani , Mehdi, Consulting advisor
Translated Title Supplied by Cataloguer : تعیین شرایط بهینه تولید نانوبادی اختصاصی مولکول تنظیم کننده ایمنی TIM-3 با استفاده از تکنیک نمایش فاژی
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Determination of Optimized Condition for Production of Ca...
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