Establishment Of Double Antibody Sandwich Elisa For Mouse IL-18 And IL-10

Interleukin(IL)is a class of cytokines produced by various cells,which is vital for regulating immune response and inflammation.Therefore,establishment for an immunoassay of IL is of great significance.The study is aiming at establishing double antibody sandwich ELISA methods with high sensitivity and good specificity for the detection of mouse IL-18 and IL-10.The results are described as follows:(1)The mouse IL-18 protein was obtained from the Escherichia coli expression system.Six monoclonal antibodies,including 4G4,3G11,4C4,3E8,4E6 and 4A10,were obtained by monoclonal antibody preparation technique.We selected m Ab 4E6 as the capture antibody and 4G4 as the detection antibody to establish a double-antibody sandwich ELISA for the detection of mouse IL-18 after antibody pairing test.The experimental results showed that the limit of detection was 17.7 pg/m L(2?)and the linear range of the method was 40-1400 pg/m L.The detection method did not show any cross-reactivity with mouse IL-10,IL-1?,IL-17 A,IL-2,IL-6 and IFN-?,demonstrating a good specificity of the method.(2)The study of chapter was based on eight hybridomas of IL-10 obtained in the preliminary work.The cross-reaction experiment showed only six hybridomas(1F3,4E3,2E3,3D8,4F9,1F9)were specific to mouse IL-10.Five monoclonal antibodies including4E3,1F3,2E3,3D8 and 4F9 were obtained by monoclonal antibody preparation technique.We selected m Ab 3D8 as the capture antibody and m Ab 4F9 as detection antibody to establish a double-antidody sandwich ELISA for the detection of mouse IL-10 after antibody pairing test.The experimental results showed that the limit of detection was 61.3ng/m L(2?)and the linear range of the method was 0.08-2.56 ?g/m L.There is a certain gap between this ELISA method and the commercial kits.