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Iranian Journal of Biotechnology، جلد ۱۱، شماره ۳، صفحات ۱۸۲-۱۹۲
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عنوان انگلیسی Cloning, Expression and Characterization of Recombinant Human Fc Receptor Like 1, 2 and 4 Molecules
چکیده انگلیسی مقاله Background: The Fc receptor like (FCRL) molecules belong to the immunoglobulin (Ig) superfamily with potentially immunoregulatory function. Among the FCRL family FCRL2 and 4 are predominantly expressed on memory B cells and FCRL1 is a pan- B cell marker. To date, no ligand has been identifid for the human FCRL1, 2 and 4 molecules. Objectives: Cloning, expression, purifiation and structural analysis of the extracellular domain of human FCRL1, 2 and 4 proteins. Materials and Methods: In this study, the extracellular part of human FCRL1, 2 and 4 were subcloned into prokaryotic expression vectors pET-28b (+) and transformed into BL21-DE3 E.coli strain. Protein expression was optimized by fie adjustments such as induction time, incubation temperature and expression hosts. Recombinant FCRL proteins were purifid by metal affity chromatography using Ni-NTA resin. Purifid FCRL proteins were further characterized by SDS-PAGE and immunoblotting using His-tag and FCRL specifi polyclonal antibodies. Results: Our results demonstrated that FCRL1, 2 and 4 were successfully expressed in pET-28b (+) vector. Optimization of the expression procedure showed that IPTG induction at OD600 = 0.9 and overnight incubation at 37˚C resulted in the highest expression levels of FCRL proteins ranging from approximately 15% (FCRL1) to 25% (FCRL2 and 4) of the total bacterial lysate proteins. Conclusions: These purifid recombinant proteins are potentially a valuable tool for investigating the immunoregulatory function of FCRL molecules and the production of specifi mAbs for immunotherapeutic interventions.
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نویسندگان مقاله مهدی شعبانی |
monoclonal antibody research center, avicenna research institute, acecr, tehran, ir iran and department of immunology, school of medicine, shiraz university of medical sciences, shiraz, ir iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی شیراز (Shiraz university of medical sciences)

اعظم همتی |
monoclonal antibody research center, avicenna research institute, acecr, tehran, ir iran

مهدی zandemami |
monoclonal antibody research center, avicenna research institute, acecr, tehran, ir iran

جلال خوشنودی |
department of immunology, school of public health, tehran university of medical sciences, tehran, ir iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی تهران (Tehran university of medical sciences)

محمود جدی تهرانی | jeddi tehrani
monoclonal antibody research center, avicenna research institute, acecr, tehran, ir iran

حجت اله ربانی |
monoclonal antibody research center, avicenna research institute, acecr, tehran, ir iran

زهرا امیرغفران |
department of immunology, school of medicine, shiraz university of medical sciences, shiraz, ir iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی شیراز (Shiraz university of medical sciences)

فاضل شکری |
monoclonal antibody research center, avicenna research institute, acecr, tehran, ir iran

نشانی اینترنتی http://www.ijbiotech.com/article_7198_49132f6c7406d1140cdc8c0dbe33e594.pdf
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