A novel method using immuno-affinity chromatography for isolating β-conglycinin from soybean proteins

A monoclonal antibody (Mab) 6G(4) against soybean beta-conglycinin has been prepared using a conjugate of chicken ovalbumin and a synthetic peptide that corresponded to one of the epitope sequences of beta-conglycinin as the immunogen. An ELISA method for the quantification of beta-conglycinin has also been developed. In the present study. we report a novel method for the purification of beta-conglycinin by Mab 6G(4)-based immuno-affinity chromatography. beta-Conglycinin with a purity of 92.9% was successfully isolated from soybean proteins. Western blot assay was used to further identify its characteristics and the results demonstrated that the purified beta-conglycinin maintains its biological activities. Therefore, the Mab-based immuno-affinity chromatography is an available method for purification of beta-conglycinin. It also provides a new opportunity for future study on the mechanism of food allergy responses using high purity beta-conglycinin as the experimental material. (C) 2009 Elsevier Ltd. All rights reserved.