Tehran University of Medical Sciences Iranian Journal of Allergy, Asthma and Immunology 1735-1502 19 2 2020 04 16 139 148 EN Mitra Hosseinpour 15 Khordad Hospital, ShahidBeheshti University of Medical Sciences, Tehran, Iran AND Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Kazem Mashayekhi Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Reza Falak Department of Immunology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran Sirous Jamalzehi Department of Medical Laboratory Sciences, Iranshahr University of Medical Sciences, Iranshahr, Iran Saeed Mohammadian Haftcheshmeh Department of Medical Immunology, Nanotechnology Research Center, Bu-Ali Research Institute, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Mohamad Javad Mousavi Bushehr University of Medical Sciences, Bushehr, Iran AND Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Anvar Soleimani Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Khadijeh Koushki Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Mojtaba Sankian Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Mohammad Soukhtanloo Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran 2019 05 26 2019 11 12 Allergy to non-specific lipidtransfer protein (nsLTP), the major allergen of grape (Vit v1), is considered as one of the most common fruit allergies in Iran. Therefore, a specific monoclonal antibody (mAb) can be used for the characterization and assessment of. Accordingly, this study aimed to generate and characterize a mAb against Vit v1 with a diagnostic purpose. To this end, Vit v1 allergen (9 kDa) was extracted using a modified Bjorksten extraction method. Natural Vit v1-immunized mouse splenocytes were fused with SP2/0Ag-14 myeloma cells for generating hybridoma cells. Specific antibody-secreting Hybridoma cells were selected using ELISA. Finally, anti-Vit v1 mAb was characterized by western blotting, ELISA, and isotyping methods. In the current study, a 9 kDa (Vit v1) protein was attained fromcrude and fresh juice of grape extracts and the isotype of desired anti-Vit v1 mAb was determined as IgM with k light chain. In addition, The ELISA results demonstrated that anti-Vit v1 mAb was specified against natural Vit v1 in the grape cultivar and related LTP allergens, such as Pla or 3 (p<0.0001). In the present study, a specific mAb was produced for detecting the LTP allergen. This mAb with a confirmed specificity can be utilized for evaluating the LTP allergens and their allergenicity in different grape cultivars. https://ijaai.tums.ac.ir/index.php/ijaai/article/view/2416