Efficient and accurate AAV capsid quantification
Efficiently and accurately quantifying the physical titre of adeno-associated viral (AAV) vectors in bioprocessing is crucial for process development, as well as maintaining consistency, efficacy, and safety of gene therapy products.
With increased pressure to standardise and optimise viral-vector production processes to achieve higher yields and lower cost of goods, rapid, accurate, at-line AAV capsid titre measurement is essential for process optimisation.
Confidently capture AAV titre results, at-line in real-time
Caught between the analytical delays of centralised equipment or the resource-heavy hands-on experience of standard ELISA for your AAV quantification?
Our Amperia Redox Electrochemical Detection (RED) system offers a cost-effective, automated alternative for efficient, accurate AAV capsid quantification. The compact Amperia instrument can be deployed conveniently at-line for rapid results that support immediate process optimisation decisions.
Automated, accurate AAV quantification at the bench or bioreactor
Streamline decision-making and optimise bioprocess efficiency; access actionable AAV capsid titre results conveniently at any bench or bioreactor site direct from unpurified samples.
- Deploy anywhere: optic-free and compact, the Amperia can be deployed wherever you need it most
- Simple-to-use: automated workflow consistency with built in analytics and no specialist training required
- Crude compatible: RED highly sensitive and specific direct detection enables AAV quantification from crude cell culture lysates across a broad range (1×109 vp/mL to 1×1011 vp/mL) saving time on purification or serial dilutions and preserving precious samples.
- Time efficient: automated processing reduces hands-on time by 96% compared to traditional ELISA and delivers results 3 x faster.
- Accurate and precise: average error rate <6% from a known concentration across replicates compared to up to 16% average error rate demonstrated by ELISA in a recent study.
- Cost-effective: re-usable calibration curves reduce the costs of reagents and AAV standards.
- Flexible throughput: Amperia uses standard 96-well microplates – just need a quick answer on a few samples, no need to wait and store samples to fill a full automated ELISA run, get your answer immediately.
Simplified AAV capsid quantification using Amperia RED
Streptavidin
Capture molecule
Antibody
Capture antibody
AAV
Sample
HRP tagged antibody
Detection reagent
STEP 1
Streptavidin
Pre coated streptavidin sensors are placed inside the instrument.
STEP 2
Capture antibody
An anti AAV capture antibody is bound to streptavidin once the sensors are dipped into the first set of wells.
STEP 3
Sample capture
AAVs in sample bind to the capture molecule and antibody complex.
STEP 4
Detection antibody
A detection antibody conjugated to HRP binds to the analyte to prepare for electrochemical analysis.
STEP 5
Electrochemical measurement
Redox Electrochemical Detection “RED” takes place, whereby the electrochemical interaction between enzyme and substrate is captured an interpreted as a concentration based off of a standard curve. Specifically the conversion of TMB into TMB+ in the presence of HRP and therefore, binded sample.
Graph showing the percentage difference (error) between the average AAV titre measurements and the true concentration for Amperia RED and ELISA methods across low and high concentrations and three days. Amperia RED shows an average error of less than 6% across all days. ELISA shows an average error of up to 16%.
Fast, accurate, reliable AAV quantification, a comparison with ELISA
In this study we compared the accuracy, reproducibility and efficiency of the Amperia RED system with traditional ELISA assay techniques when measuring AAV capsid titre across a range concentrations. Amperia showed greater accuracy and consistency, together with significantly reduced hands-on time and circa 3 x faster time to result.
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