Lumit® dsRNA Detection Assay
IVT mRNA サンプル中の二本鎖RNA（dsRNA）汚染を検出・定量するアッセイ
- 高い特異性：一本鎖RNA（ssRNA）やDNAとの交差反応なし
- 定量性：配列やサイズに依存せず（30bp以上に対応）
- シンプルなワークフロー：洗浄ステップや固相化なしで実施可能
- 新規アプローチ：dsRNAの検出に抗体を使用しない方法を採用
Catalog Number:
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Size
Each assay kit includes its respective dsRNA standard. If you need extra standard, select the option from the dropdown to add additional vials.
Catalog Number: W2041
Catalog Number: W2042
Catalog Number: W2051
Catalog Number: W2052
Catalog Number: W2061
Catalog Number: W2062
Catalog Number: W2071
Catalog Number: W2072
Catalog Number: W2081
Catalog Number: W2082
Catalog Number: W2010
Catalog Number: W2040
Catalog Number: W2050
Catalog Number: W2060
Catalog Number: W2070
Catalog Number: W2080
Quantify dsRNA with Confidence
High-quality in vitro transcription (IVT) products are crucial for research and RNA therapeutic development. However, IVT can generate double-stranded RNA (dsRNA) byproducts. This contaminant can have significant negative downstream effects on drug substances and products, such as high immunogenicity. Sensitive and quantitative assessment of dsRNA is critical to control and optimize mRNA therapeutic and vaccine manufacturing processes.
The Lumit® dsRNA Detection Assay is a novel technique to quantify dsRNA in a sample. Lumit® Assay technology is based on NanoBiT® luciferase complementation. NanoBiT® subunits, SmBiT and LgBiT, are genetically fused to dsRNA binding proteins. In the presence of dsRNA, the SmBiT and LgBiT subunits are brought together to form an active NanoBiT® Luciferase, generating a luminescent signal proportional to the amount of dsRNA in a sample. The result is a concentration-dependent luminescent signal for quantitative and specific measurement of dsRNA.
Note: The total number of assays per kit will vary based on final well volume.
For standard well volumes:
- 100 assay kits: applicable for 100 assays in 96-well plates or 400 assays in 384-well plates.
- 500 assay kits: applicable for 500 assays in 96-well plates or 2,000 assays in 384-well plates.
Simple Protocol Streamlines dsRNA Quantification
The assay is solution-based and homogeneous:
- No wash or sample transfer steps needed
- Amenable to high-throughput applications
Size and Sequence-Independent dsRNA Assay: The Lumit® dsRNA Detection Assay can sensitively quantify dsRNA independent of sequence and size.
Better Performance than Commercial ELISA Kits: The Lumit® dsRNA Detection Assay provides an accurate linear range of 0.04 to 2.5ng/ml. Commercial dsRNA ELISA kits do not accurately quantitate dsRNA at these concentrations.
Detect and Quantify dsRNA in Complex IVT Samples: The Lumit® dsRNA Detection Assay can accurately quantify dsRNA in IVT samples with mixed RNA species.
Detecting dsRNA
The Lumit® dsRNA Detection Assay exhibits outstanding intra- and inter-assay precision and extremely low limit of detection.
Standard response curve generated using the Lumit® dsRNA Detection Assay across a series of dsRNA concentrations. Each point represents the mean of six replicates, and the data were collected from three independent experiments. Measurements were recorded using a GloMax® Discover plate luminometer. The assay demonstrated a calculated Limit of Detection (LoD) of 0.37pg/ml and a Limit of Quantitation (LoQ) of 1.12pg/ml, highlighting its exceptional sensitivity and reproducibility.
|
Inter-Assay Precision |
|||
| Expected (ng/ml) |
Apparent n=48 (ng/ml) |
Recovery n=48 (%) |
CV (%) |
| 0.125 | 0.130 | 104 | 6.2 |
| 0.5 | 0.494 | 99 | 5.2 |
| 2.0 | 2.069 | 103 | 3.6 |
| Average: 5 | |||
Three concentrations of dsRNA were tested as replicates of 16 in three experiments. Average apparent concentrations were determined by linear interpolation using the standard curve.
Detecting Modified and Unmodified dsRNA with Lumit® dsRNA Assay
Standard curve response range of the modified dsRNA Standards. Standard curves were prepared for each modified dsRNA Standard: pseudouridine (Ψ), N1-methylpseudouridine (m1Ψ), 5-methyoxyuridine (mo5U) and 5-methylcytidine (m5C). Luminescence was plotted after subtracting background, using a GloMax® Discover System and a 0.5-second integration time. This figure is for illustration only. Always perform a standard curve on each experimental plate. Absolute RLU values will vary due to luminometer sensitivity and settings.
Specific Detection of dsRNA in IVT Products
The Lumit® dsRNA Detection Assay can accurately quantify dsRNA in IVT samples with mixed RNA species.
Lumit® dsRNA Detection Assay quantification of dsRNA in IVT samples with mixed RNA species. IVT samples were treated ± RNase If (ssRNA-specific) or RNase III (dsRNA-specific); total RNA (QuantiFluor® RNA) and dsRNA (Lumit®) were quantified.
- Left: Digestion of ssRNA with RNase If significantly reduced total RNA but did not impact the amount of dsRNA detected.
- Right: Digestion of dsRNA with RNase III into fragments ≤20bp results in a complete loss of activity in the Lumit® dsRNA Detection Assay.
See how Promega can support your therapy development — from research to production.
Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 1 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 1 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 1 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 1 × 15μl |
dsRNA Standard |
W204A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 5 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 5 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 5 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 5 × 15μl |
dsRNA Standard |
W204A | 5 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 1 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 1 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 1 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 1 × 15μl |
N1-methylpseudouridine dsRNA Standard |
W205A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 5 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 5 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 5 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 5 × 15μl |
N1-methylpseudouridine dsRNA Standard |
W205A | 5 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 1 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 1 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 1 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 1 × 15μl |
Pseudouridine dsRNA Standard |
W206A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 5 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 5 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 5 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 5 × 15μl |
Pseudouridine dsRNA Standard |
W206A | 5 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 1 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 1 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 1 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 1 × 15μl |
5-methoxyuridine dsRNA Standard |
W207A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 5 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 5 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 5 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 5 × 15μl |
5-methoxyuridine dsRNA Standard |
W207A | 5 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 1 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 1 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 1 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 1 × 15μl |
5-methylcytidine dsRNA Standard |
W208A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Lumit® Detection Substrate B |
VB405A | 5 × 160μl |
dsRNA Assay Buffer (5X) |
W201A | 5 × 8ml |
Lumit® dsRNA Sensor-LgBiT |
W202A | 5 × 15μl |
Lumit® dsRNA Sensor-SmBiT |
W203A | 5 × 15μl |
5-methylcytidine dsRNA Standard |
W208A | 5 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
dsRNA Assay Buffer (5X) |
W201A | 1 × 8ml |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
dsRNA Standard |
W204A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
N1-methylpseudouridine dsRNA Standard |
W205A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Pseudouridine dsRNA Standard |
W206A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
5-methoxyuridine dsRNA Standard |
W207A | 1 × 15μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
5-methylcytidine dsRNA Standard |
W208A | 1 × 15μl |
Resources
Featured Resource
Poster: Development of novel luminescent assays for sensitive and specific quantitation of double-stranded RNA in mRNA vaccines
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