The Lenti-X Maxi Purification Kit produces outstanding yields of highly purified virus from crude supernatants. The gravity column-based protocol is fast, simple, and effective, and superior to filter-based purification systems, which can damage fragile lentiviral particles and reduce yields. The gravity-flow column gently retains virus particles from the supernatant while unbound material flows through the column during the wash steps. Eluted virus is recovered fully intact and fully functional. Purifying virus is now as simple as purifying plasmid DNA.
Why Purify Your Lentivirus?
Lentivirus purification enables you to remove cellular contaminants that could otherwise adversely affect your transduction experiments. Crude supernatants contain residual plasmid DNA; uncharacterized infection and transduction inhibitors; cellular and serum proteins; and highly immunogenic viral proteins, nucleic acids, and virus fragments. Before using lentivirus with sensitive target cells or for in vivo applications, it is essential to remove these contaminants by purifying the lentivirus.
Our lentivirus purification columns arrive prepacked and ready to use. Simply add the 10X binding buffer to your supernatant sample (9–45 ml) and apply it to the column. The sample and buffers flow through the resin by gravity—just like a column-based plasmid prep. Impurities are removed during two column washes, and purified lentivirus is recovered in 3 ml of elution buffer.
Using purified lentivirus stocks also prevents ‘pseudo-transduction’, where high levels of recombinant protein in the crude supernatant are passively transferred to target cells during infection. Pseudo-transduction of target cells can mask the de novo expression that is expected from your transduced lentivirus.
Higher Yield and Higher Purity
A gentle purification procedure is the key to maintaining viral infectivity and producing excellent lentivirus yields of 60–80%. Anion exchange-based membrane systems offer much lower recoveries, usually less than 20%. Lenti-X columns also attain a much higher degree of virus purity than filter-based systems. In fact, purified Lenti-X samples contain very little detectable protein, while filter-purified virus preparations contain high levels of extraneous proteins that copurify with the virus.