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Lentivirus Concentrate: Virus recovery rate as high as 92%
Lentiviral vectors can infect both dividing and non-dividing cells. They offer several advantages, including a large transgene‑insertion capacity, prolonged expression of the target gene, and a low propensity to elicit host immune responses. Consequently, lentiviruses have become one of the most commonly used vector systems for expressing heterologous genes or shRNAs, with increasingly broad applications.
Introduction to Lentiviral Concentrate
EZ Lentivirus Concentration Solution is a rapid method for concentrating lentiviral particles. By incubating the un-concentrated viral supernatant with this product at low temperature and then performing low-speed centrifugation, you can obtain a concentrated viral preparation. Unlike traditional methods such as ultrafiltration or ultracentrifugation, which are time-consuming and labor-intensive, this product offers simple, fast, and safe operation with high recovery rates.
Advantages of Lentiviral Concentrate
✔ High concentration ratio: Concentration factor up to 240-fold; for example, 120 mL of viral supernatant can be concentrated down to 500 µL.
✔ High recovery rate: Testing has shown a virus recovery rate as high as 92%;
✔ High‑concentration titer: Testing has shown that, after concentration with this product, the viral titer can often reach 10⁸ or 10⁹ infectious particles.
✔ High viral activity: After concentration, the virus exhibits enhanced transduction efficiency.
Lentivirus Concentrate Operating Procedure
① Collect the lentiviral supernatants from 293T cells at 48 h and 72 h, centrifuge at 4°C and 2000 g for 10 min.
② Filter the supernatant through a 0.45 μm membrane to remove cellular debris. [Note]: Use low‑protein‑adsorbing cellulose acetate or polyethersulfone (PES) membranes. Do not use nitrocellulose (NC) membranes under any circumstances.
③ According to the volume of lentiviral supernatant: mix the concentrated reagent with the supernatant at a 4:1 ratio, shake well, and let stand at 4°C overnight, or incubate on ice for at least 3 hours. Extending the incubation time appropriately can improve the recovery rate of lentivirus.
④ After incubation, centrifuge at 4°C at 3,500 × g for 15 minutes, and carefully aspirate the supernatant.
⑤ Gently resuspend the lentiviral pellet by repeatedly pipetting up and down with an appropriate volume of PBS.
⑥ Retain a small aliquot of the viral supernatant for titer determination; aliquot the remainder and store at −80°C. [Note]: Freeze–thaw cycles can significantly reduce lentiviral titers, so please ensure that the virus is aliquoted prior to storage.
Precautions for Lentivirus Concentrate
• This product is intended solely for scientific research and must not be used for clinical diagnosis, treatment, or any other purposes.
• All lentivirus-related experimental procedures must be performed within a biosafety cabinet.
• Ensure thorough mixing of the concentrated reagent and the viral solution.
• The growth status of cells is critical for viral packaging; therefore, it is essential to maintain optimal cell culture conditions and minimize the number of passages.
• Do not subject the virus to repeated freeze–thaw cycles. If freezing and thawing are unavoidable, limit the number of cycles to no more than three, as each cycle results in approximately 10% viral loss. Aliquot the virus according to the amount needed for each use and store it at −80°C. Avoid storing it for longer than six months.
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