Production and characterization of monoclonal antibody against saffron pollen profilin, Cro s 2
IJI-Iranian Journal of Immunology. 2008; 5 (3): 156-162
in English
| IMEMR
| ID: emr-86760
ABSTRACT
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
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Index:
IMEMR (Eastern Mediterranean)
Main subject:
Pollen
/
Enzyme-Linked Immunosorbent Assay
/
Allergens
/
Crocus
/
Mice, Inbred BALB C
/
Antibodies, Monoclonal
Limits:
Animals
Language:
English
Journal:
Iran. J. Immunol.
Year:
2008
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