Recombinant human insulin analogue of the original preparation of anti-insulin monoclonal antibody immune.docVIP

 PAGE \* MERGEFORMAT 17 Recombinant human insulin analogue of the original preparation of anti-insulin monoclonal antibody immune 【Abstract】 Objective To prepare anti-insulin monoclonal antibody (mAb). Methods of recombinant human insulin analogue of the original immune BALB / c mice were prepared by immune spleen cells, the use of lymphocyte hybridoma technique with mouse myeloma cells SP2 / 0 fusion. Resulted in the creation of the seven anti-insulin mAb secreting hybridoma cell line, named A, B, C, D, E, F and G; 7 Zhu mAb respectively, of the Ig subclasses IgM, IgM, IgM, IgG2b, IgG1, IgG1 and IgG1; in the sandwich ELISA test, seven mAb are able to with human, bovine and porcine insulin-specific immune response occurs; in Western Blot tests, seven mAb are able to with the combination of recombinant human insulin analogue of the original rendering a protein blot band. Conclusion 7 mAb against human, bovine and porcine insulin were immune reactivity can be used for the determination of insulin in humans and animals and purification. Keywords: monoclonal antibody (mAb); insulin; recombinant human insulin analogue of the original Development of monoclonal antibodies against insulin by immunization with recombinant human proinsulin analog 【Abstract】 Objective To prepare monoclonal antibody (mAb) against insulinMethods Spleen cells from BALB / c mice immunized with recombinant human proinsulin analog were fused with SP2 / 0 myeloma cells by using hybridoma techniques.Results Seven hybridomas secreting mAb against insulin were established and designated A, B, C, D, E, F and G. The subclass of the seven mAb was IgM, IgM, IgM, IgG2b, IgG1, IgG1 and IgG1 respectively. The seven mAb could specifically react with insulin derived from human, cattle or pig in sandwich ELISA. The specific reaction of each mAb with recombinant human proinsulin analog was confermed by immunoblotting assay, only one stained protein band appeared on the transf