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Recombinant lentiviruses or lentiviral (LV) vectors serve as efficient vehicles in delivering transgenes to target cells in gene-modified cell therapy. Lentiviral vector titers, both total/physical and infectious, as well as vector copy number (VCN) in transduced cells, are critical quality attributes (CQAs) in viral vector manufacturing and should be optimized in process development and accurately measured in manufacturing quality control. The Applied Biosystems ViralSEQ Lentivirus Titer kits are easy-to-use, highly sensitive, and robust qPCR solutions for lentivirus characterization that enable validation per industry expectations.

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Rapid qPCR-based lentivirus quantitation

The Lentivirus Physical Titer Kit measures genome-containing lentiviral particles by RT-qPCR, and the Lentivirus Proviral DNA Titer Kit quantifies integrated proviral DNA in transduced cells for infectious titer and vector copy number assessment. These kits provide a convenient method to quantitate and correlate total and infectious LV particles, facilitating analytics in process development, optimization, and manufacturing QC.

Image of the ViralSEQ Lentivirus Testing Kit product box and tubes

Robust performance for process development and manufacturing QC

The ViralSEQ Lentivirus Physical Titer Kit offers rapid and reproducible quantitation across the production workflow; at harvest, during purification, and at lot release. With an assay range of over 7 logs, measuring and comparing LV yields across production batches is simplified. All reagents for one-step RT-qPCR and an RNA control are provided, with an optional sample preparation kit available separately.

The ViralSEQ Proviral DNA Titer Kit provides sensitive and rapid quantitation of integrated proviral DNA copies in transduced cells. Assay detection sensitivity of 10 copies and limit of quantitation (LOQ) of 25 copies allow copy number assessments in cells with low transduction efficiencies. Proviral copy numbers can be used to calculate infectious titer for lentiviral vectors or vector copy number for transduced cells. All reagents for qPCR and a DNA control are provided, with an optional sample preparation kit available separately. 

Features of ViralSEQ Lentivirus Titer Kits:

  • Seamless qPCR-to-qPCR comparison of genomic (total) and proviral (infectious) copy numbers
  • LTR-based platform assay design compatible with more than 200 transfer plasmids
  • TaqMan-assay based detection for specific quantitation
  • Rapid, actionable results in less than 6 hours
  • Optimized workflow from sample preparation to data analysis
  • Designed to meet validation criteria per industry standards

Highly specific to lentiviral RNA and proviral DNA target

Assay specificity is critical for the validation of qPCR-based analytical method. Taqman-Assay based ViralSEQ Lentivirus Titer assays show specific target amplification, and no cross-reactivity against potential contaminants that may be found in a bioproduction environment.

Cross-reactivity comparison of ViralSEQ Lentivirus Titer Kit versus SYBR Assay

Figure 2: A cross-reactant panel containing potential DNA contaminants (genomic and plasmid) common to bioproduction environment was generated to test assay specificity. Both ViralSEQ Lentivirus Titer assays show no cross-reactivity. Commercially available SYBR-based RT-qPCR (physical titer) and qPCR (proviral DNA titer) showed cross-reactivity against several species in the panel.


Streamlined workflow solution—results in less than 5 hours

ViralSEQ Lentivirus Titer Kits provide a robust and convenient method to quantitate and correlate total and infectious titers using a platform approach. These kits fit seamlessly into the broader sample-to-results workflow solution to further simplify your analytical development of LV vector based therapeutics or LV manufacturing.

We provide a complete assay workflow that includes sample preparation optimized with PrepSEQ Nucleic Acid Sample Prep Kit, for manual or automated extraction on Pharma KingFisher Flex system, with real-time qPCR on Applied Biosystems QuantStudio 5 or 7500 Fast system and data analysis using AccuSEQ(TM) Software.

ViralSEQ workflow process represented by four images of the KingFisher Flex, qPCR products, QuantStudio 5, and AccuSEQ Software.

Get consistent performance regardless of sample matrix complexity

PrepSEQ kits for residual host cell DNA sample preparation

Our Applied Biosystems PrepSEQ Nucleic Acid Sample Preparation Kit is a magnetic bead-based nucleic extraction and purification system optimized for DNA and RNA recovery from bioproduction samples. The kit provides superior recovery efficiency for applications where consistent absolute quantitation and high sensitivity detection are required.


Increase sample processing throughput using automated sample preparation

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The Applied Biosystems Pharma KingFisher Flex 96 Deep-Well Magnetic Particle Processor offers an easy workflow with high throughput so you can process up to 96 samples with 1 hour of total hands-on time using PrepSEQ sample preparation kits.

  • Consistent, high-quality nucleic acid recovery from a wide variety of sample types
  • Validated nucleic acid purification method workflow to enable consistent, reproducible results
  • Cross-contamination control leveraging the bead transfer technology

Simplify data analysis with integrated instrument and analysis software

The Applied Biosystems AccuSEQ Real-Time PCR Detection Software supports the needs of analytical testing for viral vector characterization during the biopharmaceutical manufacturing process, as well as routine qPCR assays. The software is designed to work with the Applied Biosystems QuantStudio 5 (QS5) Real-Time PCR Instrument with 0.1 mL, 96-well plates and the Applied Biosystems 7500 FAST Real-Time PCR instrument.

Key features:

  • Interfaces with the 7500 FAST and QS5 Fast Real-Time PCR instruments
  • Intuitive interface to set up sample runs
  • Security, audit, and e-Signature capabilities to help enable 21 CFR 11 compliance
  • Windows 10 compatible

Additional analytical solutions for viral vector-based therapeutics

Save further analytical development resources with additional Applied Biosciences molecular-based assays designed to support required analytics for both biopharmaceutical development, in-process, and manufacturing lot release testing.  These include residual host cell DNA and residual plasmid DNA quantitation, rapid real-time PCR detection of mycoplasma contamination, and genotypic microbial identification for environmental monitoring.

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