Abstract: To establish hybridoma cell lines that secreted antiGly m Bd 28K monoclonal antibodies， Gly m Bd 28K protein was used to immunize BALB/c mouse， whose spleen was removed for cell fusion. Two hybridoma cell lines that stably secreted antiGly m Bd 28K monoclonal antibodies were screened and then were injected into BALB/c mice to prepare monoclonal antibodies. After purification of monoclonal antibody， two strains of cell lines were identified for IgG1， named as mAb1E3 and mAb2F4. Indirect ELISA was used to detect the immunological properties of mAb1E3. The titer of mAb1E3 was 1∶4 10×105， and the IC50 of mAb1E3 to Gly m Bd 28K was 23.99 μg·L-1. The ratios of crossreactivity of mAb1E3 with peanut protein， sesame protein， walnut protein and wheat glutenin were less than 0.1%. We successfully prepared the antiGly m Bd 28K monoclonal antibodies possessed high sensitivity and specificity. This study provided a basis for the detection of Gly m Bd 28K allergen and the identification of epitopes.

Key words: Gly m Bd 28K, hybridoma cell, monoclonal antibody, indirect ELISA