A
monoclonal antibody (Mab), 4B2, against soybean
glycinin was prepared using the preliminary extracted natural
glycinin as the immunogen in our previous study. Herein, we established a novel method for the purification of
glycinin by Mab 4B2-based immunoaffinity chromatography. The characteristics of the purified
glycinin were identified by SDS-PAGE, Western blot, and histamine release assay.
Glycinin was successfully isolated from soybeans with a yield of 16.8% and a purity of 93.8%, which were significantly higher than those produced using other traditional procedures. The acidic
polypeptides of the purified
glycinin can be recognized by the Mab 4B2, but not the basic
polypeptides. In addition, the histamine release ratio of the purified
glycinin was similar to that of natural
glycinin, which indicated that the purified
glycinin maintained its
biological activities. Further study revealed that the Mab/gel ratios ranging from 6.0 to 12.0 mg/mL were suitable for the isolation of
glycinin using immunoaffinity chromatography. Taken together, this new method based on immunoaffinity chromatography could be used for high-yield and high-purity natural
glycinin production and would facilitate future study on the mechanism of soybean-induced
food allergy.