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" Optimizing the selection and production process of ssDNA aptamer against α4 integrin "
\ Shirin Abdollah Kouhpayeh Isfahani
; Abbas Rezaei, Hossein Khanahmad
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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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109355
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| Doc. No
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T17322
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QU57,A135o,2016
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| Main Entry
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Abdollah Kouhpayeh Isfahani, Shirin
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| Title & Author
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Optimizing the selection and production process of ssDNA aptamer against α4 integrin\ Shirin Abdollah Kouhpayeh Isfahani
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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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xi, 80p.: ill, diag, tab
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| Note
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This is a research dissertation with project ID: 392607
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شیرین عبداله کوهپایه اصفهانی
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| Abstract
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Multiple sclerosis (MS) is considered as an autoimmune demyelinating disease of central nervous system (CNS). The pathogenesis of MS gives a clue for scientists to target the important molecules involved in this process. One of the most important molecules for MS pathogenesis is α4 integrin which is responsible for autoreactive leukocytes migration into the brain. The monoclonal antibody, Natalizumab, was introduced to market for blocking the extravasation of autoreactive leukocytes via inhibition of α4 integrin. However, the disadvantages of antibodies provided a suitable background for other agents to be replaced with antibodies. Considering the profound advantages of aptamers over antibodies, aptamer isolation against α4 integrin was intended in the present study. The α4integrin-specific aptamers were selected using cell-SELEX (Systemic Evoultion of Ligands by EXponential Enrichment) method with Human Embryonic Kidney (HEK)-293 overexpressing α4 integrin and HEK-293 as target and control cells, respectively. Evaluation of selected aptamer was performed through flow cytometric analysis. The selected clones were then sequenced and analyzed for possible secondary structure and affinity. The results of this study led to isolation of 13 different ssDNA clones in 11 rounds of selection which were categorized to three clusters based on common structural motifs. The evaluation of SELEX progress showed growth in aptamer affinity with increasing of the number of cycles. Taken together, the findings of this study demonstrated the isolation of α4-specific ssDNA aptamers with suitable affinity for ligand, which can further be replaced with Natalizumab.
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| Descriptor
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Aptamers, Nucleotide
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Multiple Sclerosis
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Oligonucleotides
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Integrin alpha4
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| Added Entry
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Rezaei, Abbas, Thesis advisor
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Khanahmad, Hossein, Thesis advisor
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| Translated Title Supplied by Cataloguer
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بهینه سازی مراحل گزینش و تولید آپتامر SSDNA ضد آلفا 4 اینتگرین
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