Development of a monoclonal antibody-based competitive ELISA for detection of beta-conglycinin, an allergen from soybean

Soybean beta-conalycinin is one of the major food allergies for children and young animals. In order to detect immunoreactive beta-conglycinin from soybean and soybean products, monoclonal antibodies against beta-conglycinin were prepared using a conjugated chicken ovalbumin with a synthetic peptide that corresponded to one epitope sequence of beta-conglycinin as the immunogen. The generated monoclonal antibodies. named as 6G(4), 3B(7), and 5F(10), were identified as being of IgG(1) isotype and exhibited high specificity to beta-conglycinin. A competitive enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibody 6G(4) was established to determine P-conglycinin and showed an IC50 value of 4.7 ng/mL with a detection limit of 2.0 ng/mL. The recovery tests of P-conglycinin indicated that the monoclonal antibody 6G(4)-based competitive ELISA gave reliable reproducibility. Therefore, the produced monoclonal antibody 6G(4) and the developed competitive ELISA could provide a valuable tool for sensitive determination of P-conglycinin and for future studies on food allergies related to soybean P-conglycinin. (c) 2007 Elsevier Ltd. All rights reserved.