International Journal of Medical Laboratory

en ی A Mouse Monoclonal Antibody Against Human IFN-γ and its Characters ایمنی شناسی Immunology پژوهشي Research ی <div style="text-align: justify;">Background and Aims: A monoclonal antibody (mAb) can unambiguously identify, quantify, and purify an antigen or particular epitope at a large scale. The superiority of these antibodies lies in their specificity for the antigenic determinant. So, this study aims to prepare mouse mAb-secreting hybridoma against human gamma interferon (IFN-γ) and determine the produced antibody's characters. Materials and Methods: Mouse splenic B lymphocytes immunized with recombinant human IFN-γ were fused with mouse SP2/0 cells. The hybridized cells were selected by hypoxanthine-aminopterin-thymidine and hypoxanthine-thymidine media to obtain monoclonal antibody-producing hybridoma cells. Finally, indirect enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and western blot were used to confirm the creation of antibody-secreting hybridoma cells. Results: mAb against IFN-γ were produced by fusing SP2/0 mouse non-secretory myeloma cell line with the spleen cells of immunized mice. This antibody's indirect ELISA optical density was 2.055 on average, and the desired antibody bands were confirmed in SDS-PAGE compared to Septicol® (commercial antibody). Also, in the western blot, the desired antibody could bind to the antigen. IFN-γ transferred on nitrocellulose membrane. In ELISA and western blot tests, anti-mouse IgG conjugated antibodies were used; therefore, the mAb IgG isotype was taken into consideration. Conclusion: In this study, a mouse mAb was obtained by immunization of Balb/C mice and fusion of spleen cells of these mice with the SP2/0 cells, which can specifically bind to recombinant human IFN-γ and can be used to detect IFN-γ secretion in all types of intracellular infections, including latent tuberculosis.</div> Hybridoma, IFN-γ, Monoclonal antibody, SP2/0 67 74 http://ijml.ssu.ac.ir/browse.php?a_code=A-10-212-10&slc_lang=en&sid=1 Erfan Zaker عرفان ذاکر fatemezare91@gmail.com Yes Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Department of Genetics and Molecular Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Department of Genetics and Molecular Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Fateme Zare فاطمه زارع fatemezare91@yahoo.com 0000-0003-0436-9924 No Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Seyed Hossein Hejazi سیدحسین حجازی ijml.office@gmail.com No Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Hossein Khanahmad حسین خان احمد hossein_khanahmad@yahoo.com No Department of Genetics and Molecular Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Department of Genetics and Molecular Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Seyed Mehdi Kalantar سیدمهدی کلانتر smkalantar@yahoo.com 0000-0002-6994-6449 No Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran