4.2 Article

Immunological Characterization of Polyclonal Antisera Prepared Against Recombinant Rice RAG2 and Its Application in Detection of 14-16 kDa α-amylase/trypsin Inhibitors from Processed Foods

Journal

FOOD SCIENCE AND TECHNOLOGY RESEARCH
Volume 16, Issue 6, Pages 599-606

Publisher

KARGER
DOI: 10.3136/fstr.16.599

Keywords

rice (Oryza sativa); RAG2; alpha-amylase/trypsin inhibitor; recombinant protein; polyclonal antisera; processed food

Funding

  1. Ministry of Agriculture, Forestry and Fisheries of Japan

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RAG2 is a member of the rice 14-16 kDa alpha-amylase/trypsin inhibitors, which are the main allergens in rice grains. Recombinant RAG2 protein was expressed in Escherichia coli, and polyclonal antisera were prepared against the protein. The polyclonal antisera specifically bound with 14-16 kDa alpha-amylase/trypsin inhibitors from husked brown, red and black rice. No cross-reaction was observed between the polyclonal antisera and saline-soluble proteins from other tested cereal grains. By immunoblotting with the polyclonal antisera, 14-16 kDa alpha-amylase/trypsin inhibitors were found to be distributed throughout the endosperm of brown rice and their contents were estimated. Moreover, 14-16 kDa alpha-amylase/trypsin inhibitors were also detected in processed foods. Thus, the polyclonal antisera prepared against recombinant RAG2 protein could be used to compare the content of 14-16 kDa alpha-amylase/trypsin inhibitors in rice samples and probe the presence of the allergen in processed foods.

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