A one-step RT-qPCR assay for the quantitation of genome-containing lentiviral particles in supernatants from cell-based bioproduction systems. Lentiviral quantitation is a critical quality attribute (CQA) in viral vector manufacturing for cell and gene-modified cell therapy development.
The Applied Biosystems ViralSEQ Lentivirus Physical Titer Kit is a one-step RT-qPCR assay for the quantitation of genome-containing lentiviral particles in supernatants from cell-based bioproduction systems. Lentiviral quantitation is a critical quality attribute (CQA) in viral vector manufacturing for cell and gene-modified cell therapy development.
ViralSEQ Lentivirus Physical Titer Kit is an RT-qPCR assay for the quantitation of genome-containing lentiviral particles in supernatants from cell-based bioproduction systems. The assay targets a conserved region (LTR) in the lentiviral vector genome and can be used with most lentiviral transfer plasmids. Compared to p24 ELISA methods, lentiviral quantitation by RT-qPCR is more reproducible and specific to genome-containing particles.
Features of the ViralSEQ Lentivirus Physical Titer Kit: • Lentiviral genome specific RT-qPCR assay for accurate viral particle count • Assay design compatible with more than 200 lentivirus transfer plasmids • Includes master mix, TaqMan assay mix and RNA control • Configuration with PrepSEQ kit available for optimized RNA recovery
Integrated workflow solution The ViralSEQ Lentivirus Physical 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 ViralSEQ kit has been validated on the Applied Biosystems 7500 Fast Real-Time PCR System and QuantStudio 5 Real-Time PCR System. Data analysis is streamlined using AccuSEQ Real-Time PCR Detection Software that provides accurate quantitation and security, audit, and e-signature capabilities, to help enable 21 CFR Pt 11 compliance.
Broad dynamic range The kit includes a synthetic RNA control to generate a standard curve, as well as internal positive controls. It enables quantitation over 7 logs of dynamic range from 50 copies to 109 copies for a range of titer values. TaqMan Assay-based detection allows for highly specific quantitation.
Genomic detection and quantitation of viral particles Achieve specific quantitation of genome-containing lentiviral particles using an RT-qPCR assay. Methods relying on measurement of HIV-1 p24 could overestimate viral particle count to include partial capsids and free p24. The ViralSEQ Lentivirus Physical Titer kit measures viral particles based on viral RNA, thereby improving quantitation specificity of genome-containing particles.
Complete lentivirus characterization ViralSEQ Lentivirus Physical Titer Kit can be used with the ViralSEQ™ Lentivirus Proviral DNA Titer Kit (Cat. No. A53561) 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.
Specifications
Detection MethodPrimer-probe
For Use With (Equipment)Real-time PCR instruments (QuantStudio5 or 7500 Fast)