production and characterization of monoclonal antibody against saffron pollen profilin, cro s 2

background: allergy to saffron (crocus sativus) pollen has been described in people involved in processing of saffron flower stamens. profilins have been identified as a pan-allergen in different plant pollens and foods. this molecule is an actin-binding pro-tein with a molecular weight of 12-16 kda found in eukaryotic species. objective: the aim of this study was to generate monoclonal antibody against cro s 2 in order to char-acterize this major allergen of saffron pollen. methods: balb/c mice were immunized to obtain adequate humoral response. splenocytes were prepared from the immunized animals, mixed with the p3-x63-ag8.653 myeloma cells and fused by means of peg 1500. after two weeks of culturing in hat-containing media, the supernatant from those wells growing hybridomas were screened by elisa using plates coated with cro s 2. cells from positive wells were cloned at least 3 times by limiting dilution. specific-ity and cross-reactivity of the mabs were determined by western blot analysis and sandwich elisa. results: two stable hybridoma clones secreting mabs against cro s 2 were obtained and expanded. the anti-cro s 2 mabs were also found to cross-react with other plant profilins. isotype of this mab was identified as μ heavy chain and k light chain. conclusion: the anti-cro s 2 mab could be a useful tool for characteriza-tion and standardization of many pollen and fruit-derived profilins.