Centrifugation of 14000 rpm for 30 min was no so efficient. After size analysis there are still present liposomes in the supernatant. Liposomes are composed of mixture of lecithin, dotap dope and chol. All your helps are appritiated.
Hi there
The Ghosh Research Group (UC Riverside) uses 60,000 g for 1 to 2 hours to concentrate liposomes of 100nm.
Gel filtration would also be possible as well. Perhaps you could focus on that and size-exclusion chromatography (SEC) in general instead of on centrifugation? There are several studies using SEC to study different exosomes in body fluids. I think it wouldn't be a bad idea to look some of them up.
I hope I could help
Art
It's difficult to thoroughly pellet some types of liposomes because of their low buoyant density.
One alternative is to use ultrafiltration, either with a centrifugal ultrafiltration unit or a stirred pressure cell. A disadvantage of this method, however, is that the liposomes tend to form a gel against the membrane, which might affect the size distribution.
Another way that might work is to put the liposome sample in dialysis tubing, then reduce the volume by immersing the dialysis tubing (or cassette) in an absorbent, such as polyacrylamide beads (e.g. Bio-gel) or Sephadex G-100, occasionally brushing off the swelled adsorbent and exposing the tubing to fresh unswollen absorbent. This will take several hours, but it is very easy to do.
- Badges
- Science topic
- Similar topics
- Methodology
- Laboratory