Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA
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Stojanović, Marijana M.
Jankov, Ratko M.
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Thaumatin-like proteins (TLPs) have been established as a new family of fruit and pollen allergens. The aim of this study was to develop a two-site ELISA for the quantification of the thaumatin-like kiwi allergen (Act d 2) in kiwifruit extracts and kiwi fruit-containing food products. Genomic DNA (gDNA) of Act d 2 was amplified and the deduced amino acid sequence was determined to obtain a primary structure. Act d 2 purified from kiwifruit extract by HPLC was identified by Edman degradation and MS. Balb/c mice were immunized with Act d 2 for the production of mAbs by hybridoma technology. The optimized ELISA measured Act d 2 concentrations ranging from 0.2 to 9.0 ng/mL, with intra- and interassay coefficients of variation of 3.65 and 10.44%, respectively. The developed ELISA is a useful method for the quantification of the thaumatin-like kiwi allergen in kiwifruit extracts as well as the allergen level in kiwifruit-containing food products. It may be a helpful analytical tool for the e...valuation of the stability (integrity) of fruit allergen extracts for in vitro diagnosis.
Keywords:Act d 2 / food allergy / immunoassay / kiwifruit / thaumatin-like protein
Source:Molecular Nutrition and Food Research, 2008, 52, 6, 701-707
- Wiley-Blackwell, Malden