Determination of Pea Proteinase Inhibitors Using ELISA Based on Monoclonal Antibodies |
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| Authors: | Anne Marie Arentoft Hanne Fr⊘kiær Hilmer S⊘rensen Susanne S⊘rensen |
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| Affiliation: | 1. Chemistry Department , Royal Veterinary and Agricultural University , 40 Thorvaldsensvej, DK-1871, Frederiksberg C, Denmark;2. Department of Biochemistry and Nutrition , The Technical University of Denmark , Building 224, DK-2800, Lyngby, Denmark |
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| Abstract: | Abstract An immunoassay method for analysis of trypsin and chymotrypsin inhibitor activity in seeds from different pea cultivars has been developed. This method is fast, cheap and well suited for screening of large numbers of seed samples. The technique is based on enzyme linked immunoassay (ELISA) using monoclonal antibodies produced against pea (Pisum sativum L.) proteintype trypsin inhibitors (PPI). The results obtained by ELISA have been compared with results achieved by using traditional enzymatic analyses for determination of both trypsin and chymotrypsin inhibitor contents, and these methods showed good agreement. Great differences in PPI levels have been found between various pea cultivars grown in Finland and Denmark, and these differences comprised both the total amount of PPI and the relative composition of individual PPI. Several proteins occurring in pea exhibited inhibitor activity, and at least 10 PPI inhibitors with p/ values 4.9–7.8 were detected. Pea cultivars with low PPI levels had a PPI composition different from the PPI composition found in cultivars with high PPI levels. |
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| Keywords: | Pisum trypsin inhibitors chymotrypsin inhibitors enzyme linked immunoassay mAbs |
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