Lentiviral vectors have a broad host range, enabling efficient infection of both dividing and non-dividing cells. They integrate into the host cell genome, thereby achieving stable functional expression or silencing of genes in various cell types. This provides possibilities for cell labeling, gene function research, target discovery, and cell and gene therapy.
The manufacturing of lentiviral vectors involves several steps: production, purification, storage, and gene transfer. Lentiviruses are extremely fragile enveloped viral vectors—factors such as shear force, extrusion, temperature, pH, ionic strength, and operation time during production can significantly affect their activity. Additionally, lentiviruses are prone to aggregation, which poses considerable challenges to their production and storage.Typically, lentiviral vectors are stored at -80°C after filling. For use, they need to undergo freeze-thaw cycles and infect target cells at 37°C. At low temperatures, proteins may tend to denature, and lipid bilayers may easily lose structural integrity. Lentiviral vectors usually require long-term storage at -80°C because repeated freeze-thaw cycles of the viral vector can lead to loss of transduction ability. However, lentiviral vectors exhibit poor stability when stored in conventional physiological buffers; their activity decreases rapidly during storage at -80°C or repeated freeze-thaw cycles. This increases the cost of lentiviral vectors and limits their applications. Therefore, a protective solution capable of stably storing viral vector formulations at low temperatures is particularly important.
Aladdin has developed a Lentivirus Stable Storage Solution (L1505998) that enhances the viability of lentiviral vectors and ensures their stability. Using this product to resuspend lentiviral particles can improve the stability of lentiviral vectors at -80°C and during repeated freeze-thaw cycles, maintain the activity of lentiviral vectors, and enhance the transduction efficiency of lentiviral vectors in target cells. Meanwhile, the reagents used in this product have well-defined chemical compositions, which facilitate industrial-scale production and application.
Product Features:
⁕ High viral activity.
⁕ Maintains the stability of lentiviral vectors during storage at -80°C or repeated freeze-thaw cycles.
⁕ Prevents viral aggregation and enhances transduction capability.
⁕ Well-defined chemical composition, facilitating industrial-scale production and application.
Experimental Steps:
1. Transfer the medium containing lentiviral particles to a sterile container, and centrifuge at 800×g for 10 minutes to remove cell debris.
⁕ To ensure viral activity, the viral supernatant before concentration should preferably be freshly collected undiluted viral solution.
2. Filter the supernatant through a 0.45 μm filter into a sterile container.
⁕ If using a membrane filter, low protein-binding cellulose acetate or polyethersulfone (PES) membranes are recommended; nitrocellulose (NC) membranes are not recommended.
3. Add 1 volume of Lentivirus Concentration Reagent (5×, S1506239) to every 4 volumes of lentivirus-containing supernatant, diluting the Lentivirus Concentration Reagent (5×) to the working concentration (1×).
4. Incubate the above mixture slowly in a shaker at 4°C for 3 hours or overnight.
⁕ Lentiviral particles can remain stable at 4°C for up to 4 days.
5. Centrifuge the mixture at 4000×g and 4°C for 25 minutes. After centrifugation, lentiviral particles may appear as a beige or white precipitate at the bottom of the container.
6. Carefully discard the supernatant, then centrifuge again at 4000×g and 4°C for 5 minutes. Discard all residual liquid thoroughly, taking care to avoid aspirating the precipitate—this precipitate is the lentiviral particles.
7. Resuspend the viral particles with this product (the Lentivirus Stable Storage Solution) at a volume of 1/10 to 1/100 of the initial volume (original supernatant volume). Gently pipette up and down to resuspend the lentiviral particles.
8. Aliquot the lentivirus into small volumes, store them in a -80°C refrigerator, and reserve a small amount of virus for titer determination.
⁕ Repeated freeze-thaw cycles of the lentivirus should be avoided as much as possible, as this will reduce the viral titer.
Scope of Application:
Suitable for the low-temperature stable storage of any lentiviral particles.
Precautions:
1. For your health and safety, please operate in compliance with standard procedures and wear a lab coat and gloves when conducting experiments.
2. All lentivirus-related operations must be performed in a BSL-2 (Biosafety Level 2) biological safety cabinet.
3. This product is for research use only and shall not be used for clinical diagnosis or treatment.
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