Clipos™ NTA Liposomes, Non PEGylated

Catalog:CDEIMS-1512

Unit Size:2 mL, 5 mL

INQUIRY Datasheet

Specifications
Description It is well established that electron-rich ligands such as histidine, tryptophan or cysteine show a relatively high affinity to bond with electropositive transition metals, including Co+2, Ni+2, Cu+2 and Zn+2. This observation has been exploited to improve and control the association of diverse histidine-tagged peptides to liposomes containing metal-chelating lipids. Therefore, immunoliposomes can be generated using nickel-chelating lipids such as Ni-nitrilotriacetic acid (NTA) and 1,2-dioleoyl-sn-glucero-3-[N-(amino-1-carboxypentyl)-iminodiacetic acid) succinyl] (DOGS-NTA aka DGS-NTA), which are commercially available, and His-tagged proteins or peptides. When the liposomes with Ni-NTA headgroups are combined with the His residues, typically at the N- or C-terminus of proteins, the proteins reversibly anchor to the liposomes.
Lipid composition L-a-PC/Cholesterol/1,2-Dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (nickel salt) (69/30/1 molar ratio)
Liposome Size 100 nm
Lipid Mass Concentration 14.84 mg/mL
Lipid Molar Concentration 22.45 mM
Functional Group NTA
Appearance 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.
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 4 months.
Technical Notes Higher concentration of DOGS-Ni-NTA lipid in the liposome formulation will cause a increased change of nonspecific binding and therefore it is important to keep the molar percentage of chelating DOGS-Ni-NTA lipid below 3% to avoid nonspecific binding. Immunosome®-Ni-NTA and Immunodox®Ni-NTA kits all contain 1% DOGS-Ni-NTA which is an optimized concentration for specific binding to His6-tagged proteins, peptides and antibody fragments.
The binding reaction between a His6-tagged peptide and DOGS-NTA-Ni liposomes will be inhibited in the presence of high concentration of imidazole (~166 mM).
It should be noted that the use of a 10-histidine tag promoted liposome–liposome crosslinking in the absence of PEG-modified lipids. This could be explained by suggesting that the stretch of 10 histidine residues would be sufficient to bind more than one DOGS-NTA-Ni liposome.
It has been demonstrated that the presence of PEG polymers inhibits surface reactions between liposomes, and thus can be used to prevent aggregation.
If you are using a ligand or peptide that is hydrophobic then 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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