Clipos™ DiA labeled Liposomes, Cyanur Lipid, PEGylated
Specifications
| Description
|
Proteins can be covalently coupled to the liposomes via amine-reactive cyanur-groups, either directly to the vesicle surface using cyanuric chloride-activated DSPE (cyanur-DSPE) or to the distal ends of PEG-spacers using activated cyanur-PEG-PE (ammonium salt). Cyanuric chloride at the PEG terminus functions to link peptides, antibodies and other amine-containing biomolecules or nanoparticles via a nucleophilic substitution reaction under basic conditions. Antibodies or other proteins can be conjugated without any previous derivatization. |
| Lipid composition
|
HSPC/Cholesterol/DSPE-PEG(2000)/DSPE-PEG(2000)-Cyanur (~57/38/4/1 molar ratio) |
| Liposome Size
|
100 nm |
| Lipid Mass Concentration
|
~15.92 mg/mL |
| Lipid Molar Concentration
|
~21.58 mM |
| Functional Group
|
Cyanur |
| Fluorophore
|
DiA |
| Excitation
|
456 nm |
| Emission
|
590 nm |
| Appearance
|
Be colored and the color is depend on the type of the fluorescent dye that is used. 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. |
| Buffer
|
Phosphate Buffered Saline, pH 7.4 |
| Storage
|
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. |
| Shelf-life
|
2 months. |
| Technical Notes
|
Tris buffer should never be used in any step of the process since it contains amine.
Cyanuric chloride is considered as a sensory respiratory irritant. However, despite the name of the cyanur-modified liposomes, they have not shown any sign of acute, chronic or genotoxicity.
Cyanur groups are amine-reactive, however, some random attachments of the antibodies can be expected since cyanuric chloride can react with a wide range of nucleophilic functionalities, such as alcohols and thiols. This may interfere with the binding of the antibody to the liposome, and therefore, the binding affinity would change.
Size exclusion spin columns such as Sepharose® CL-4B can be used instead of Float-A-Lyzer® dialysis cassette. However a very large amount of liposomes will stick to the column during the cleanup process and therefore we strongly suggest using dialysis than size exclusive beads.
If you are using a ligand or peptide that is hydrophobic, it is recommended to solubilize it in DMSO or DMF and then add the buffer to it. It is recommended not to use more than 5% volume of DMSO or DMF in the solution. DMF and DMSO are both compatible with liposomes and they are also miscible in water. Other organic solvent such as ethanol and chloroform are not compatible with liposomes and will cause the liposomes to lyse. If you end up using DMSO or DMF then after the conjugation reaction is done, you need to remove DMSO and DMF from the liposomes. In order to do that you need to use a dialysis cassette that is made from REGENERATED CELLULOSE MEMBRANE. NOTE: Not all membranes are compatible with DMF and DMSO. We recommend using a Slide-A-Lyzer™ MINI Dialysis Device with MWCO of 2K made from regenerated cellulose membrane manufactured by ThermoFisher. After DMSO or DMF is removed, you can use Float-A-Lyzer® dialysis device for the final step of cleaning up the prep. |
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Liposomes Production
Datasheet