Monoclonal Antibodies Raised Against Wheat HMW-GS And Antigenic Determinant Prediction

The compositions of high molecular weight glutenin subunits (HMW-GS) are closely related to bread-making quality of wheat flour. The routine measurement of bread-making quality needs expensive apparatus and larger amount of samples which are generally insufficient at the early stage of breeding program. Immunological method with the advantages of lower cost, smaller sample amount, more simple handling, has been utilized to predict and screen wheat quality. Monoclonal antibodies (McAb) are generated by single clone of B cell after stimulating of corresponding antigen. McAbs raised against wheat glutenins appear to be powerful tool in the studies of wheat storage proteins and screening of wheat quality. HMW-GS 1Bx17 and lByl8 have been confirmed to have positive effects on wheat bread-making quality, and therefore it is necessary to prepare McAbs specific to this two subunits. Until now, the lByl8 gene has not been separated by the traditional PCR method. McAb specific to lByl8 as probes to screen wheat cDNA expressing libraries could provide us alternative way to clone wheat storage protein genes.In this study, HMW-GS lBxl7+lByl8 was separated and identified from bread wheat cultivar Kontrast by SDS-PAGE and RP-HPLC, then purified by gel electrophoresis method. The purified components were applied as antigen to immune BALB/c mice. McAb were prepared by traditional hybridoma method. Applications of the McAbs on bread-making quality were primarily studied.Nine hybridomas were prepared, named as 1E1F7C5F7D4, 1E1F7C5F7E7, 1E1F7C5F7F4, 2C7G6D4G4E5, 2C7G6D4G4F2, 2C7G6D4G4H6, 1E1F7C5F7B11, 1E1F7C5F7E4 and 1E1F7C5F7E2.In addition, bioinformatic methods were used to predict epitopes from HMW-GS. The results obtained could be helpful for further study.