TruTiter® Reagent System
Assess the integrity of AAV capsids for consistent quantification of viral genomes using dPCR or qPCR
- Consistent quantification of AAV genome
- No DNase treatment required
- Simple and fast workflow
- Membrane-impermeable to maintain viral integrity
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Catalog Number: A8884
Catalog Number: A8885
TruTiter® Reagent: AAV Quantification You Can Trust
Adeno-associated virus (AAV) is a commonly used vector for gene therapy delivery. One of the main challenges for AAV gene therapy is developing effective quality control and quality assurance protocols, because accurate quantification and characterization of AAV production batches is essential for proper clinical dosing. Current protocols, such as DNase treatment and mammalian infectivity assays, are often inadequate and can lead to a high degree of lot-to-lot variability, which can result in non-optimal trial outcomes due to inaccurate determination of functional gene dosage.
Contaminating DNA can interfere with accurate AAV genome titer determination prior to qPCR or dPCR. DNase treatment to eliminate contaminating DNA is often cumbersome and lacks the precision needed for accurate determination of viral titer due to inconsistent digestion.
TruTiter® Reagent System enables consistent quantification of AAV viral genome without the DNase treatment step.
TruTiter® Technology Principle
- TruTiter® Reagent System uses a novel molecule that is membrane-impermeable and can bind to any exposed nucleic acids, preventing any downstream amplification, thus eliminating the DNase treatment step in the AAV genome titer workflow.
- The TruTiter® Reagent is compatible with existing PCR viral quantification protocols, allowing direct amplification using a variety of qPCR or digital PCR platforms.
With TruTiter® Reagent, only the DNA inside the intact capsid can be amplified for accurate genome titer.
TruTiter® Reagent System
Simple and Fast Assay Workflow
Assay Performance Data
TruTiter® Reagent Results in Less Variability
Compared to DNase-Based dPCR Workflows
AAV2-NanoLuc® viral capsids (1010GU/ml) were treated at 65°C for 10 minutes. Following heat treatment, the AAV particles were treated with either DNase (red) or TruTiter® Reagent (green) and amplified using dPCR with 8 technical replicates each.
AAV2-NanoLuc® viral capsids (1010GU/ml) were treated with 0.1% Tween®-20 for 10 minutes. Following detergent treatment, the AAV particles were treated with DNase (red) or TruTiter® Reagent (green) and amplified using dPCR with 8 technical replicates each.
AAV2-NanoLuc® viral particles (107GU/ml) were treated at the indicated temperature for 10 minutes. Following temperature treatment, the AAV particles were treated with DNase (red) or TruTiter™ Reagent (green) followed by dPCR. In parallel, a 24-hour infectivity assay (blue) was performed using HEK293 cells. To aid interpretation, data is represented as percent of signal achieved with the room temperature (R.T.) treatment.
Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
Viability PCR Reagent |
AM230A | 1 × 250μl | 1mM |
Neutralization Buffer B (100X) |
AM310A | 1 × 500μl |
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Neutralization Buffer B (100X) |
AM310A | 1 × 500μl |
Resources
Featured Resource
Poster: A Novel Technology for Accurate Quantification of Viral Genome Titers
The TruTiter® Reagent is a small molecule that is excluded from intact structures such as AAV capsids. However, the molecule covalently conjugates exposed nucleic acid and prevents subsequent amplification by PCR. This property enables selective amplification of fully encapsulated nucleic acids for accurately measuring intact AAV genome titers.
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