Production and Characterization of Monoclonal Antibody against Vit v1: A Grape Allergen Belonging to Lipid Transfer Protein Family

摘要

Allergy to non-specific lipidtransfer protein (nsLTP), the major allergen of grape (Vit v1), isconsidered as one of the most common fruit allergies in Iran. Therefore, a specific monoclonalantibody (mAb) can be used for the characterization and assessment of. Accordingly, this study aimedto 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 forgenerating 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 grapeextracts and the isotype of desired anti-Vit v1 mAb was determined as IgM with κ light chain. Inaddition, The ELISA results demonstrated that anti-Vit v1 mAb was specified against natural Vit v1 inthe grape cultivar and related LTP allergens, such as Pla or 3 (p0.0001).In the present study, a specific mAb was produced for detecting the LTP allergen. This mAb witha confirmed specificity can be utilized for evaluating the LTP allergens and their allergenicity indifferent grape cultivars.