production and characterization of monoclonal antibody against vit v1: a grape allergen belonging to lipid transfer protein family

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 κ 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.