Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA
| Autor: | Milena Spasic, Ljiljana Dimitrijević, Aleksandra Inic-Kanada, Marija Gavrović-Jankulović, Tanja Cirkovic Velickovic, Arnd Petersen, Wolf-Meinhard Becker, Buko Lindner, Marijana Stojanovic, Ratko M. Jankov |
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| Rok vydání: | 2008 |
| Předmět: |
Actinidia
Molecular Sequence Data Enzyme-Linked Immunosorbent Assay medicine.disease_cause Thaumatin-like protein Mice 03 medical and health sciences 0302 clinical medicine Allergen Food allergy medicine Animals Amino Acid Sequence Cloning Molecular Plant Proteins 030304 developmental biology 2. Zero hunger Immunoassay Mice Inbred BALB C 0303 health sciences Hybridomas Chromatography biology medicine.diagnostic_test Edman degradation Plant Extracts Antibodies Monoclonal food and beverages Allergens biology.organism_classification medicine.disease genomic DNA 030228 respiratory system Thaumatin Kiwi Fruit Act d 2 Hybridoma technology Kiwifruit Food Analysis Food Science Biotechnology |
| Zdroj: | Molecular Nutrition and Food Research |
| Popis: | 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 evaluation of the stability (integrity) of fruit allergen extracts for in vitro diagnosis. |
| Databáze: | OpenAIRE |
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