Messenger RNA (mRNA) is a groundbreaking platform for vaccines and therapeutics. However, during in-vitro transcription (IVT), double-stranded RNA (dsRNA) can form as an unwanted by-product. Even trace amounts of dsRNA can trigger innate immune responses, reduce translation efficiency, and compromise product performance.

To ensure product safety and meet rising regulatory expectations, Abnova provides a dsRNA impurity analysis. This service combines qualitative detection (Dot Blot) and quantitative measurement (ELISA) to accurately assess dsRNA contamination in mRNA products, supporting research, process development, and quality assurance.

Advantages

Dot Blot
  • Rapid and Simple Workflow
  • Qualitative & Semi-Quantitative Assessment
  • Low Sample Consumption
  • Cost-Effective Analysis
 

ELISA
  • High Sensitivity & Quantitative Accuracy
  • High Throughput Capability
  • Excellent for Low-Level Detection
 
 

Our Services
  • Dot Blot
  • Enzyme-Linked Immunosorbent Assay (ELISA)
 
 

Specifications 
Service Item Description
Analyte Double-stranded RNA molecules
Methods Dot Blot (qualitative)
Sandwich ELISA (quantitative)
Reference Material poly I:C
Specificity Check Heat denaturation (95°C, 5 min)
Limit of Detection Dot Blot:  0.05 ng dsRNA in 1ug total RNA
Sandwich ELISA: 1.17 ng/mL
Sample Type IVT mRNA
Sample Requirement 10 ug, 0.1 ug/ul
Turnaround Time 5 working days
Deliverables Dot Blot image, ELISA data, calibration curve, dsRNA concentration, COA
 
 

Examples

Comparison of dsRNA Levels in Unpurified and Purified mRNA by Dot Blot and ELISA

Dot Blot
mRNA_dsRNA_Dot_Blot_ELISA_1.jpg
Sample Unpurified mRNA 1 Purified mRNA 1
Dot Blot (ng dsRNA in 1 μg total RNA) 0.32~1.25 0~0.32
 
mRNA_dsRNA_Dot_Blot_ELISA_2.jpg (674 KB)
Sample Unpurified mRNA 2 Purified mRNA 2
Dot Blot (ng dsRNA in 1 μg total RNA) 1.6~6.25 0.064~0.25
 

 ELISA
mRNA_dsRNA_Dot_Blot_ELISA_3.jpg (618 KB)
Sample Unpurified mRNA 1 Purified mRNA 1
dsRNA (ng/mL) N.D. N.D.

N.D. means that the limit of detection of this assay is lower than 1.17 ng/mL.
 
mRNA_dsRNA_Dot_Blot_ELISA_4.jpg (618 KB)
Sample Unpurified mRNA 2 Purified mRNA 2
dsRNA (ng/mL) 6.176 N.D.

N.D. means that the limit of detection of this assay is lower than 1.17 ng/mL.
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