Double-stranded RNA (dsRNA) emerges as a by-product during the in vitro transcription of mRNA. This by-product possesses immunogenic properties within the human body, capable of provoking an immune response, consequently diminishing mRNA levels. This renders dsRNA a troublesome process impurity necessitating thorough elimination and stringent control of its residual presence. To address this concern, the Double-stranded RNA (dsRNA) ELISA detection kit employs the experimental principles of a double-antibody sandwich enzyme-linked immunoassay (ELISA) for the quantification of residual dsRNA. The Kit can not only be used in the detection of regular dsRNAs, but also in detection of peudo UTP, N1-Me-peudo UTP and 5-OMe-UTP modified dsRNAs. The procedure involves adding both a standard and the test sample onto an enzyme plate (designated as 36717-A) previously coated with anti-dsRNA antibodies. Subsequently, biotin-labeled dsRNA detection antibodies (36717-F) are added in a diluted form. Finally, Streptavidin-HRP (SA-HRP) (36717-G) is introduced to create a complex comprising antibodies, antigens, biotin, and SA-HRP. Following plate washing, the addition of TMB chromogenic solution (36717-K) initiates color development. Under the influence of HRP enzyme catalysis, TMB undergoes a transformation from colorless to blue, which is then halted through the application of the stop solution (36717-L). Ultimately, this transformation results in a yellow hue, the intensity of which correlates positively with the quantity of dsRNA identified in the sample. This kit serves a versatile purpose, enabling optimization of the biological product purification process, impurity control during intermediate stages, and release testing for final product assessment. Validation Report Features High Sensitivity: Quantitative lower limit (LLOQ) reaches 0.0156 pg/μL; detection limit (LLOD) ≤0.001 pg/μL. High Precision: Intra-assay CV
