Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA

被引:17
|
作者
Gavrovic-Jankulovic, Marija [1 ]
Spasic, Milena [1 ]
Velickovic, Tanja Cirkovic [1 ]
Stojanovic, Marijana [2 ]
Inic-Kanada, Aleksandra [2 ]
Dimitrijevic, Ljiliana [2 ]
Lindner, Buko [3 ]
Petersen, Arnd [4 ]
Becker, Wolf-Meinhard [4 ]
Jankov, Ratko M. [1 ]
机构
[1] Univ Belgrade, Fac Chem, Dept Biochem, Belgrade 11000, Serbia
[2] Inst Immunol & Virol, Dept Immunol, Belgrade, Serbia
[3] Borstel Res Ctr, Div Immuno Chem, Borstel, Germany
[4] Borstel Res Ctr, Div Mol & Clin Allergol, Borstel, Germany
关键词
Act d 2; food allergy; immunoassay; kiwifruit; thaumatin-like protein;
D O I
10.1002/mnfr.200700286
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
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.
引用
收藏
页码:701 / 707
页数:7
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