The Applied Biosystems ViralSEQ Lentivirus Proviral DNA Titer Kit is a qPCR assay for the quantitation of integrated proviral DNA in transduced cells. Proviral copy number can be used to calculate infectious lentiviral titers (TU/mL) and subsequently, the volume of vector supernatant needed to transduce cells at the desired multiplicity of infection (MOI). The assay can also be used to measure vector copy number (VCN), a critical quality attribute (CQA) in cell and gene-modified cell therapy development.
ViralSEQ Lentivirus Proviral DNA Titer Kit is a qPCR assay for the quantitation of integrated proviral DNA in cells transduced with lentiviral vectors. The assay targets a conserved region (LTR) in the provirus and can be used with most lentiviral transfer plasmids. It provides a wide dynamic range and high specificity for the accurate detection of provirus copies. Proviral copy numbers can be used to calculate infectious viral titer and vector copy number in gene-modified cell therapy development.
Features of the ViralSEQ Lentivirus Proviral DNA Titer Kit:
• Lentiviral genome-specific qPCR assay for provirus quantitation
• Assay design compatible with more than 200 lentivirus transfer plasmids
• Includes master mix, TaqMan assay mix, and DNA control
• Configuration with PrepSEQ kit available for optimized DNA recovery
Integrated workflow solution
The ViralSEQ Lentivirus Proviral DNA Titer Kit is part of an integrated workflow for viral vector characterization during biopharmaceutical manufacturing. The kit is compatible with the PrepSEQ Nucleic Acid Sample Preparation Kit (Cat. No. A50485), which offers both a manual and automated sample preparation workflow. For real-time PCR, the kit has been validated on the Applied Biosystems 7500 Fast Real-Time PCR System and the QuantStudio 5 Real-Time PCR System. Data analysis is streamlined using the AccuSEQ Real-Time PCR Detection Software. The software provides accurate quantitation and security, audit, and e-signature capabilities, to help enable 21 CFR Part 11 compliance.
Sensitive detection
With detection sensitivity of 10 copies and limit of quantitation (LOQ) of 25 copies, the assay allows measurement of low proviral copy numbers. Data can be correlated to cell number to calculate VCN or to viral vector sample volume used for transduction to calculate infectious titers (TU/mL).
Broad dynamic range
The kit includes a synthetic DNA control to generate a standard curve, as well as internal positive controls. It enables quantitation from 25 genome copies to 107 genome copies per reaction. TaqMan Assay-based detection provides highly specific quantitation.
Complete lentivirus characterization
ViralSEQ Lentivirus Proviral DNA Titer Kit can be used with the ViralSEQTM Lentivirus Physical Titer Kit (Cat. No. A52597) for seamless correlation of qPCR data post-harvest (genome-containing particle count) and post-transduction (vector copy number analysis and infectious particle count). These kits provide a convenient method to correlate total and infectious lentiviral particles, facilitating lentiviral vector analytics in process development, optimization, and manufacturing QC.
For Research Use Only. Not for use in diagnostic procedures. TaqMan is a trademark of Roche Molecular Systems, Inc., used under permission and license.