초고속원심분리보다 훨씬 간단한 방법
Lenti-X Concentrator와 초고속원심분리를 이용한 바이러스 농축 방법을 비교하면 Lenti-X 법의 장점이 확연히 들어난다 (Table I). Lenti-X Concentrator 는 초고속원심분리법보다 더욱 유연하고 빠르며, 쉬울 뿐만 아니라 매우 효과적이다. 바이러스 상층액의 부피를 줄이거나 역가를 증가시켜야 할 때, Lenti-X Concentrator는 번거롭고 시간이 많이 소요되는 초고속원심분리 과정 없이 빠르고 간단하게 농축시킬 수 있다.
Figure 1. The Lenti-X Concentrator protocol.
Add Lenti-X Concentrator reagent to clarified viral supernatant, incubate for 30 min to overnight at 4°C, and spin. That’s it.
| Table I: Lenti-X Concentrator vs. Ultracentrifugation | ||
| Feature | Lenti-X Concentrator | Ultracentrifugation |
| Easily Scalable | Yes | No |
| Specialized Equipment | No | Yes |
| Time Required | ~1 hr | 4 hr to overnight |
| Ease-of-Use | ++++ | + |
| Yield | ＞90% | ＞90% |
Figure 2. Efficient concentration with minimal loss.
Lentiviral supernatant from a pLVX-ZsGreen1 vector was concentrated from 3 ml down to 30μl using the Lenti-X Concentrator Reagent and reflects a 100-fold increase in viral titer. The total amount of virus contained in each sample is also indicated, which shows that the resuspended pellet captured 90% of the virus present in original sample. Samples were titrated on HT1080 cells and analyzed by flow cytometry 48 hr post-transduction.
