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New - Biopharma Quality Control : AAV Vector Titer Determination

AAV Vector Titer Determination


Determination of adeno-associated virus vector titer by digital PCR with MiQuant® AAV.

  • Fast and easy workflow
  • Quantitation independent of AAV serotype
  • No DNA extraction required
  • Designed for QIAcuity platform
  • In-Process Control and Release Testing
  • Quantiation without standard curve or further calculations
  • Lyophilized kit components simplify logistics and storage

Gene therapy represents one of the most ground-breaking advancements in modern medicine that has rapidly evolved over the last years leading to improved methods of applications. Adeno-associated virus (AAV) vectors are among the most prominent viral vectors for in vivo gene therapy that target the healthy gene material to the correct location in the organism which enables the mutated gene to be exchanged by the healthy one.
Viral genome titer is a critical quality attribute in AAV characterization for safe and efficacious gene therapy products. Specificity of AAV vectors as well as the concentration of the vector titer must be determined throughout the production process and for final release testing.
The MiQuant® AAV kits have been developed for the determination of AAV vector titer. Different kit versions enable the specific detection of AAV genotype 2/6 (including genotype 1, 3 & 7) or genotype 5.

 

Application Areas

Determination of the AAV vector titer with the MiQuant® AAV kits is useful at different times. During the AAV vector production process, it can be used as an in-process control to evaluate the AAV titer. As part of the final QC release testing the AAV titer must be determined prior to administration to the patient. Besides these applications the MiQuant® AAV kits can also be used for determining the AAV titer in research and development.

Find more information on Minerva Biolabs' MiQuant® AAV in our flyer.

MiQuant® AAV ITR 5 – dPCR

The MiQuant® AAV ITR 5 – dPCR kit is an easy to use, rapid and sensitive solution for adeno-associated virus (AAV) vector titer testing based on digital PCR (dPCR).

Minerva Biolabs

Catalogue No.DescriptionPack SizePriceQty
58-0601MiQuant® AAV ITR 5 – dPCR35 ul primer probe mix POA Quantity Add to Order
58-0602MiQuant® AAV ITR 5 – dPCR140 µl primer/probe mix POA Quantity Add to Order

Description

MiQuant® AAV ITR 5 – dPCR


The MiQuant® AAV ITR 5 – dPCR kit is an easy to use, rapid and sensitive solution for adeno-associated virus (AAV) vector titer tes


ting based on digital PCR (dPCR).

This kit provides a primer/probe mix that enables specific detection of AAV genotype 5 by amplifying the respective inverted terminal repeat (ITR) region and is compatible with various dPCR and ddPCR devices (QIAcuity®, ddPCR™, Quantstudio™ Absolute Q™).

The integrated TruTiter™ technology (powered by Promega) ensures that only the DNA packaged within intact AAV capsids is amplified, while free DNA and DNA from broken capsids remain masked and undetectable. This enables highly precise measurement of AAV vector genome titer and eliminates the need for a DNase treatment step.

Type of PCR

Digital PCR (dPCR and ddPCR™)

Recommended Use

AAV vector titer testing of AAV ITR 5 for in-process control and final release testing.

Kit Components
  • MiQuant® AAV 5 Mix (lyophilized)
  • Positive Control DNA (lyophilized)
  • Internal Control DNA (lyophilized)
  • TruTiter™ Reagent
  • Neutralization Buffer
  • dPCR grade Water
Package Size

Cat. No. 58-0601: 35 µl primer/probe mix

Cat. No. 58-0602: 140 µl primer/probe mix

Since every dPCR device requires different PCR reaction volumes, the number of PCR reactions are shown in the following table.

Number of PCR reactions

Result Evaluation

Absolute Quantification

Required Lab Device and Consumables
  • Digital PCR device with corresponding consumables (plates or stripes)
    • QIAcuity®
    • Quantstudio™ Absolute Q™
    • QX600™ or QX200™ ddPCR™
  • Corresponding Mastermix (QIAcuity® Probe PCR Kit (4x), Absolute Q™ DNA Digital PCR Master Mix (5x), ddPCR™ Multiplex Supermix (4x) or ddPCR™ Supermix for Probes (2x))
  • Thermal shaker for temperatures up to 95°C
  • 1.5 ml reaction tubes (DNA-/RNA-/DNase-/RNase-free; preferably low-bind)
  • Microcentrifuge for 1.5 ml PCR tubes
  • Vortex mixer
  • Pipettes with corresponding filter tips

Optional:

  • Restriction enzyme MspI (20 U/µl)
  • MiQuant® SP Beads (sample preparation kit, Cat. No. 56-4010/-4100)
  • PCR Clean™ (Cat. No. 15-2025 and 15-2001)
Shipment, Shelf Life, and Storage

The Shipment is refrigerated. The kit components must be stored at ≤ -18°C until use. The rehydrated components must be stored at ≤ -18°C for a maximum of one month. The expiry date of the unopened package is marked on the package label.

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