SignaGen provides high-quality lentiviral purification using sucrose ultracentrifugation, a well-established method that yields concentrated, functional lentiviral particles while preserving infectivity. This approach is particularly suitable for research-grade and preclinical applications requiring reliable performance and consistent quality.

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Purification by Sucrose Ultracentrifugation

Following lentiviral packaging in producer cells, viral supernatants are harvested, clarified to remove cell debris, and subjected to ultracentrifugation through a defined sucrose cushion. During high-speed centrifugation, intact lentiviral particles sediment through the sucrose layer and form a concentrated viral pellet, while soluble proteins, media components, and low-molecular-weight impurities remain in the supernatant.

The viral pellet is carefully resuspended in an appropriate formulation buffer to generate a concentrated lentiviral preparation. This method is widely used due to its simplicity, reproducibility, and ability to maintain viral integrity without exposing particles to harsh chemical conditions.

Key advantages include:

• Effective concentration of lentiviral particles
• Preservation of viral infectivity
• Removal of soluble proteins and media contaminants
• Compatibility with a broad range of lentivector designs

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Quality Control (QC) Testing

Each purified lentiviral preparation undergoes a panel of quality control assays designed to confirm identity, purity, potency, and overall consistency.

1. Functional Titer Determination

Lentiviral titer is measured using a functional transduction assay in permissive cells. Titers are reported as transducing units per milliliter (TU/mL), providing a biologically relevant measure of viral potency.

2. Physical Particle Assessment

Where requested, physical particle levels may be evaluated by p24 ELISA to quantify total lentiviral capsid protein and assess particle recovery following purification.

3. Sterility Testing

Purified lentivirus is tested for microbial contamination to ensure the absence of bacterial or fungal growth.

4. Mycoplasma Detection

Mycoplasma testing is performed using sensitive molecular-based assays to confirm the preparation is free of mycoplasma contamination.

5. Endotoxin Testing

Endotoxin levels can be determined using LAL-based assays, particularly important for in vivo or sensitive cell culture applications.

6. Replication-Competent Lentivirus (RCL) Determination

Amplify from a permissive cell line for 2~3 weeks, followed by p24 readout

7. Identity Confirmation (Optional)

Vector identity is confirmed based on production records and plasmid verification data associated with the lentivector used for packaging.

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Reliable, Research-Ready Lentivirus

By combining sucrose ultracentrifugation with a comprehensive QC framework, SignaGen delivers lentiviral vectors that are consistent, potent, and suitable for demanding downstream applications. Our purification and testing workflows are designed to support reproducibility and confidence in every experiment.

For additional details on lentiviral purification options, QC specifications, or custom requirements, please contact us at info@signagen.com