Establishment Of Double Antibody Sandwich ELISA For Mouse IL-17 And Preparation Of Colloidal Gold Test Strip For Cry1Ab

In basic scientific research and practical application,various detection technologies are very important and widely used.They have their own outstanding advantages and suitable scenarios.In this study,two detection methods were constructed for two kinds of objects:1)A double antibody sandwich enzyme-linked immunosorbent assay(ELISA)with high sensitivity and specificity for quantitative detection of mouse interleukin-17(mIL-17)was established,which can be used in the study of inflammatory diseases related to mIL-17.The specific conclusions are as follows:mIL-17 protein was obtained by E.coli expression system.Seven mouse monoclonal antibodies(Mc Abs)(1E4,2B2,2D8,3A9,3C4,5F6,5G1)and rabbit polyclonal antibodies(Pc Abs)were obtained by immunizing mice and rabbits respectively.Then,we established a Mc Abs and Pc Abs sandwich ELISA which using 3C4 and Pc Abs as capture antibody and detection antibody respectively,and horseradish peroxidase(HRP)-labeled goat anti-rabbit Ig G for signal amplification.The limit of detection was 9.80 pg/m L(S/N=2),and the range of detection was 15.60–1000pg/m L(R~2=0.9817).This method has the advantages of good specificity(there was no cross reaction with mouse IL-1?,IL-6,IL-12p40,IL-18,GM-CSF and TNF-?)and reproducibility(intra-and inter-assay coefficient of variation was 0.46%–7.04%and0.34%–7.74%,respectively),and can meet the requirements of low concentration mIL-17(10 pg/m L)detection.2)A colloidal gold strip method for rapid and qualitative detection of insecticidal protein Cry1Ab in transgenic crops was developed,which can be used for rapid detection of transgenic crops in the field.The specific conclusions are as follows:mice were immunized with Cry1Ab protein and cell fusion techniques were used to obtain two monoclonal antibodies against different epitopes of Cry1Ab,1F11 and 3H6,respectively.A colloidal gold strip detection system was designed,in which gold labeled 3H6 was used as capture antibody,while 1F11 and goat anti-mouse Ig G were respectively fixed on test(T)line and control(C)line.Cry1Ab can be detected within 15 min by this method,and the detection limit was 60 ng/m L.