Cellsome® made from Anionic PEGylated Lipids
Liposome formulations can be broadly divided into cationic, anionic and neutral subtypes. Charged liposomes are used in many different types of studies such as, blood complement studies, gene transfer studies and other biomedical applications to name a few. Cellsome® liposomes made from anionic PEGylated lipids provide certain benefits.
Anionic liposomes offer a number of attributes, relative to the challenges associated with cationic formulations such as, the administration of cationic liposomes results in higher cytotoxicity levels, both in vitro (Zhong et al., 2009) and in vivo (Balazs and Godbey, 2010). Anionic liposomes demonstrate greater stability in solution (Zhigaltsev et al., 2002, Phillips and Heydari, 1996), lead to lower aggregation when compared with neutrally charged liposomes, and have increased endocytosis when compared with cationic (Bajoria et al., 1997) and neutral liposomes.
To improve liposome stability and enhance their circulation times in the blood, a sterically-stabilized, hydrophilic polymer, polyethylene glycol (PEG), has been shown to be the optimal choice for obtaining sterically-stabilized liposomes. Using an Anionic Liposome with PEG will protect the liposomes from circulating proteins, improving their plasma clearance and enhancing their therapeutic effects.
The four products provided that are made from Anionic PEGylated Lipids provided here consist of either DSPC:Chol:DSPG:DSPE-PEG(2000) (60:30:5:5 molar ratio), DOPC:Chol:DOPG:DOPE-PEG(2000) (60:30:5:5 molar ratio), DSPC:Chol:DSPS:DSPE-PEG(2000) (60:30:5:5 molar ratio), or DOPC:Chol:DOPS:DOPE-PEG(2000) (60:30:5:5 molar ratio).
Cellsome® made from Anionic PEGylated Lipids
For more information on the lipid composition of the liposomes mentioned above click here.
|Buffer and Liposome Size||Specification|
|Buffer||Phosphate Buffered Saline*|
|Liposome size||100 nm|
|* If you prefer different buffers, please specify in your order.|
Liposome Particle Calculator
Cellsome® liposomes made from anionic PEGylated lipids are unilamellar and sized to 100 nm. The molar concentration of liposome is 10 mM. By having the liposome diameter (nm) and lipid concentration (µM), you can calculate the total number of the lipids in one liposome and number of the liposomes in one milliliter of the liposome solution. To use the calculator click here.
Each 100-nm liposome is composed of almost 80,000 phospholipid molecules. 10 mM liposome solution (phospholipid concentration of 10 mM) has 75.2 trillion (75.2×1012) liposome particles per milliliter of the solution. When using the liposomes calculator, you need to keep two things in mind:
- Enter the concentration in µM. As an example, 10 mM solution is 10,000 µM.
- Only include the concentration of phospholipids in your calculation. Cholesterol does not count since it inserts itself into the hydrophobic fatty acid chains of the lipid.
This calculation only works for the unilamellar liposomes which are below 200 nm in size. It does not work for larger multilamellar liposomes. For more information about the calculation see the video below:
- Liposomes are unilamellar and sized to 100 nm. If you need them to be made in another size, then you should mention that at the time of ordering. Unilamellar liposomes can be made in 50 nm and 200 nm sizes. Liposomes that are larger than 200 nm are multilamellar structure.
- Liposomes are made in PBS buffer at pH 7.4 but it can be made in any other buffer of your choice. You can specify your buffer at the time of ordering.
- Liposomes are made in degassed buffer that is purged with argon to avoid oxidation of the unsaturated phospholipids.
- Liposomes are made under sterile condition.
- Liposomes are unilamellar and therefore due to their small size they will not settle in the bottom of the vial.
- If you need to take multiple aliquots out of the vial, for use over a period of time, then it is advised to take extreme care in not contaminating the vial. It is recommended to handle the vial under a sterile hood to maintain the sterility of the product.
- Liposomes should never be frozen. Ice crystals that form during freezing will rupture the lipid membrane of the liposomes and change the size the liposomes particles.
Cellsome® made from anionic PEGylated lipids is a white translucent liquid made of nano size unilamellar liposomes. Usually due to the small size of liposomes no settling will occur in the bottom of the vial. The liposomes are packaged in an amber vial.
- All liposome based formulations are shipped on blue ice at 4°C in insulated packages using overnight shipping or international express shipping.
- Liposomes should NEVER be frozen. Ice crystals that form in the lipid membrane can rupture the membrane and change the size of the liposomes. Liposomes in liquid form should always be kept in the refrigerator.
- Clients who order from outside of the United States of America are responsible for their government import taxes and customs paperwork. Encapsula NanoSciences is NOT responsible for importation fees to countries outside of the United States of America.
- We strongly encourage the clients in Japan, Korea, Taiwan and China to order via a distributor. Tough customs clearance regulations in these countries will cause delay in custom clearance of these perishable formulations if ordered directly through us. Distributors can easily clear the packages from customs. To see the list of the distributors click here.
- Clients ordering from universities and research institutes in Australia should keep in mind that the liposome formulations are made from synthetic material and the formulations do not require a “permit to import quarantine material”. Liposomes are NOT biological products.
- If you would like your institute’s FedEx or DHL account to be charged for shipping, then please provide the account number at the time of ordering.
- Encapsula NanoSciences has no control over delays due to inclement weather or customs clearance delays. You will receive a FedEx or DHL tracking number once your order is confirmed. Contact FedEx or DHL in advance and make sure that the paperwork for customs is done on time. All subsequent shipping inquiries should be directed to Federal Express or DHL.
Storage and Shelf Life
Cellsome® products should always be stored at in the dark at 4°C, except when brought to room temperature for brief periods prior to animal dosing. DO NOT FREEZE. If the suspension is frozen, the encapsulated drug can be released from the liposomes thus limiting its effectiveness. In addition, the size of the liposomes will also change upon freezing and thawing.
Cellsome® made from anionic PEGylated lipids is made on daily basis. The batch that is shipped is manufactured on the same day. It is advised to use the products within 4 months of the manufacturing date.
References and background reading
1. Papageorgiou F, Pippa N, Naziris N, Demetzos C. Physicochemical Study of the Protein-Liposome Interactions: Influence of Liposome Composition and Concentration on Protein Binding. Journal of liposome research. 2018 Apr 23(just-accepted):1-29.
2. Chonn A, Semple SC, Cullis PR. Association of blood proteins with large unilamellar liposomes in vivo. Relation to circulation lifetimes. Journal of Biological Chemistry. 1992 Sep 15;267(26):18759-65.
3. Semple SC, Chonn A, Cullis PR. Interactions of liposomes and lipid-based carrier systems with blood proteins: Relation to clearance behaviour in vivo. Advanced drug delivery reviews. 1998 Jun 8;32(1-2):3-17.
4. Devine DV, Wong K, Serrano K, Chonn A, Cullis PR. Liposome—complement interactions in rat serum: implications for liposome survival studies. Biochimica et Biophysica Acta (BBA)-Biomembranes. 1994 Apr 20;1191(1):43-51.
5. Szebeni J. The interaction of liposomes with the complement system. Critical Reviews™ in Therapeutic Drug Carrier Systems. 1998;15(1).
6. Szebeni J, Baranyi L, Savay S, Milosevits J, Bodo M, Bunger R, Alving CR. The interaction of liposomes with the complement system: in vitro and in vivo assays. InMethods in enzymology 2003 Jan 1 (Vol. 373, pp. 136-154). Academic Press.
7. Bonté F, Juliano RL. Interactions of liposomes with serum proteins. Chemistry and physics of lipids. 1986 Jun 1;40(2-4):359-72.
8. Chonn A, Semple SC, Cullis PR. Association of blood proteins with large unilamellar liposomes in vivo. Relation to circulation lifetimes. Journal of Biological Chemistry. 1992 Sep 15;267(26):18759-65.
9. Colletier JP, Chaize B, Winterhalter M, Fournier D. Protein encapsulation in liposomes: efficiency depends on interactions between protein and phospholipid bilayer. BMC biotechnology. 2002 Dec;2(1):9.
10. Gregoriadis G, editor. Liposome technology: interactions of liposomes with the biological milieu. CRC press; 2006 Sep 12.
11. Papahadjopoulos D, Kimelberg HK. Phospholipid vesicles (liposomes) as models for biological membranes: their properties and interactions with cholesterol and proteins. Progress in surface science. 1974 Jan 1;4:141-232.
12. Kimelberg HK. Protein-liposome interactions and their relevance to the structure and function of cell membranes. Molecular and cellular biochemistry. 1976 Feb 1;10(3):171-90.