Production and characterization of monoclonal antibody against Saffron pollen profilin, Cro s 2.
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 protein with a molecular weight of 12-16 kDa found in eukaryotic species.



The aim of this study was to generate monoclonal antibody against Cro s 2 in order to characterize this major allergen of saffron pollen.



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. Specificity and cross-reactivity of the mAbs were determined by Western blot analysis and sandwich ELISA.



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 micro heavy chain and k light chain.



The anti-Cro s 2 mAb could be a useful tool for characterization and standardization of many pollen and fruit-derived profilins.
Sankian
Mojtaba
M
Immunobiochemistry lab, Immunology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
Glosaz-Shirazi
Forough
F
Arafi
Majid
M
Moghadam
Malihe
M
Varasteh
Abdolreza
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D027622:Crocus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D015151:Immunoblotting; D051379:Mice; D011058:Pollen / Q000276:immunology; D051304:Profilins / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D051304:Profilins