dsRNA ELISA Kit

Overview Features Product List Assay Principle Validation Data Resources Related Products

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Overview

Double-stranded RNA (dsRNA), formed from nucleic acid chains with complementary sequences, is an undesirable byproduct of in vitro transcription (IVT) during mRNA synthesis. Effective minimization and removal of dsRNA are essential to ensure the quality and safety of IVT-derived mRNA.

To detect dsRNA, the U.S. Pharmacopeia (USP) recommends methods such as dot blot and ELISA. Leveraging its advanced antibody platform, GenScript has developed a proprietary dual J2 antibody sandwich ELISA kit specifically designed for dsRNA detection. This kit offers a rapid assay time and high sensitivity, accurately detecting dsRNA species of varying lengths and chemical modifications—providing researchers with a powerful and efficient tool for quality control in mRNA production.

ELISA Kits

Key Features

Dual J2 Antibody Technology

J2 as both capturing antibody and detection antibody

Wide Linear Range

0.123-30 ng/mL

Fast Protocol

< 3 hours

Various standards

400 bp 1mψ dsRNA standard included or more options per request

Product List

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Assay Principle

dsRNA ELISA Kit

Figure 1. Schematic diagram of dsRNA ELISA with dual J2 antibodies

Validation Data

Standard Curve with Wide Linear range and High Sensitivity

Figure 2. Standard curve of ds RNA ELISA Kit (Cat. No. L01020) developed with 400 bp m1Ψ modified dsRNA standard. Linear range was detected as 0.123 – 40 ng/mL, sensitivity at 0.123 ng/mL, R² > 0.99.
Versatile dsRNA Detection

Figure 3. 3 different modification of dsRNA samples coded by same DNA and 3 dsRNA samples with different length were analyzed by dsRNA ELISA Kit (Cat. No. L01020). Result shows the all the dsRNA samples with various modification and length can be well detected.

High Accuracy

Quality Control Sample	dsRNA (ng/mL)	Intra-assay (n=10)			Inter-assay (n=30)
Quality Control Sample	dsRNA (ng/mL)	Measured dsRNA (ng/mL)	CV%	Accuracy%	Measured dsRNA (ng/mL)	CV%	Accuracy%
HQC	22.5	22.10	5.5	98.20	22.68	6.7	100.8
MQC	2.25	2.36	6.1	105.0	2.24	6.4	99.7
LQC	0.369	0.38	7.9	103.4	0.367	9.3	99.4

Table 1. Representative data of Coefficient of Variation (CV) and accuracy analyses on HQC, MQC and LQC samples from 10 runs of each sample.

Figure 4. Average of dsRNA detected on HQC, MQC and LOQC samples using 3 different batches of dsRNA ELISA Kit (Cat.No. L01020).

Wide Linear Range

Dilution Factor	Expected dsRNA (ng/mL)	Measured dsRNA (ng/mL)	CV%	Accuracy%
1000	0.738	0.755	1.5	102.3
2000	0.369	0.369	6.8	100.0
100	0.738	0.787	1.4	106.6
50	0.738	0.799	10.0	108.3

Table 2. dsRNA sample was diluted with 4 different ratios and analyzed using dsRNA ELISA Kit (Cat.No. L01020). High accuracy and low coefficient variance were presented in all testing.

High Specificity

dsRNA (ng/mL)	mRNA (ng/mL)	Measured dsRNA (ng/mL)	CV%	Accuracy%
30	300	27.79	2.2	92.6
30	30	28.21	5.4	94.0
0.123	300	0.138	4.9	112.2
0.123	30	0.115	2.5	93.5

Table 3. Two dsRNA sample (ULOQ at 30 ng/mL and LLOQ at 0.123 ng/mL) were spiked with different amount of mRNA (30ng/mL and 300ng/mL) and then analyzed with dsRNA ELISA Kit (Cat.No. L01020). High accuracy and low coefficient variance were presented in all testing.

No Hook Effect

dsRNA (ng/mL)	Absorbance (OD 450 nm)			CV%
dsRNA (ng/mL)	Duplicate 1	Duplicate 2	Average	CV%
738	4.070	4.050	4.060	0.4
73.8	3.214	3.292	3.253	1.7
36.9	2.454	2.557	2.506	2.9

Table 4. Three different dsRNA sample at various concentrations were analyzed using dsRNA ELISA Kit. No Hook Effect was observed for the high concentration sample.