Preparation and Application of Monoclonal Antibodies Against Allergen Glb- 1

BALB /c mice were immunized with G1b- 1 protein,the main allergen from wheat. The splenocytes of the immunized mice were fused with NS- 1 myeloma cells. Cell fusion and limited dilution assay were used to screen hybridoma with stable secretion character and mice ascites were induced by the selected bybridoma. Monoclonal antibodies( McABs) were purified using affinity chromatography and further tested by their specificity,subtype,titers and cross-reactivity. The antigen epitope specificity of the McABs was examined by ELISA method. Four cell lines secreting McABs against wheat G1b- 1 protein were obtained,named 1C4,4H5,1A9 and 4F5,respectively. The four McABs belong to IgG1 subtype and their titer was above 9- 10. Moreover,all the four McABs specially recognized wheat G1b-1 protein and had no cross-reaction with other familiar foods. In order to establish a double-antibody sandwich ELISA system to detect wheat GIB1 proteins,we paired off the four monoclonal antibodies for ELISA assay and found 1C4 and 4h5 might have different epitopes. Four mice anti-wheat G1b- 1 protein McABs were prepared successfully and a double-antibody sandwich ELISA method was set up in this work,which may facilitate other wheat allergen protein detection.