Double-stranded RNA (dsRNA) ELISA Kit

dsRNA is a byproduct produced during the in vitro transcription of mRNA. It has strong immunogenicity and can easily induce an immune response in the body, thereby reducing the efficacy of mRNA biological products. Therefore, dsRNA is considered a process impurity that needs to be removed during the production process and its residual amount must be strictly controlled.

This kit adopts the principle of double antibody sandwich method and combines with Biotin-Streptavidin (B-SA) amplification detection system, which is specially used for quantitative detection of double-stranded RNA (dsRNA) (≥60 bp) in samples, regardless of the sequence of dsRNA. The detection process is as follows: first, the anti-dsRNA capture antibody is pre-coated in the microwells of the ELISA plate, and the sample to be tested is added and incubated to allow the dsRNA in the sample to specifically bind to the capture antibody. After washing to remove unbound substances, the biotin-labeled detection antibody is added for incubation to form a sandwich complex of "capture antibody-dsRNA-detection antibody". After washing again, horseradish peroxidase (HRP)-labeled streptavidin (SA) is added to achieve signal amplification through the high affinity binding of biotin-streptavidin. After thorough washing, the TMB substrate is added for color reaction. TMB appears blue under the catalysis of HRP and turns yellow after the reaction is terminated by acidic stop solution. The depth of the color is positively correlated with the content of dsRNA in the sample. Finally, the absorbance (OD value) of each well is measured at a wavelength of 450 nm using a Microplate reader, and the concentration of dsRNA in the sample is calculated based on the standard curve.

This kit has high sensitivity and simple operation process, and can be widely used in the optimization of biological product purification process, the control of dsRNA impurities in the intermediate process, and the release detection of the final product.