WangLab:Magnetite Mcls

Materials

 * Chloroform
 * Bath Sonicator
 * Vortexor
 * Lipid Mix [1]
 * 1) N-(a-trimethylammonioacetyl)-didodecyl-D-glutamate chloride (TMAG)
 * 2) dilauroylphosphatidyl-choline (DLPC)
 * 3) dioleoylphosphatidylethanolamine-N-[3-(2-pyridyldithio)-propionate] (PDP-DOPE)
 * Lipid marker (?)
 * 10 nm magnetite nano-particles
 * Specific RDG peptides
 * Round bottom flask

Procedure

 * 1) Dissolve the three lipids (in a 1:2:2 molar ratio) into chloroform in the round bottom flask. Do this in a 10-20 mg of lipid/ml organic solvent ratio.
 * 2) Remove the organic solvent by either evaporating with dry nitrogen (only for volume of organic solvent less than 1ml) or through rotary evaporation.
 * 3) To make sure that all chloroform has been removed from the lipid layer, place the flask on a vacuum pump overnight. Lipid film should coat the bottom of the flask.
 * 4) Suspend the magnetite nano-particles in water.
 * 5) Hydrate dried lipid layer on the bottom of the flask with nano-particle suspended water.
 * 6) Vortex the lipid/water/nano-particle solution for a while.
 * 7) Sonicate solution for 30 mins at 28 W in a bath sonicator to reduce to single laminar liposomes.
 * 8) Add 2ml of MCLs to the RGD peptides in a .55 peptide to 1 PDP-DOPE molar ratio.
 * 9) Gently agitate this solution for 3.5 hours in a solution of 2-morpholinoethanesulfonic acid (MES) buffer (pH 5.0).
 * 10) Purify the MCLs.

Links

 * [1] Ito 2005 RDG peptide.pdf Outlines procedure for production of RGD MCLs.
 * [2] Liposome Formation.pdf General information regarding liposome formation.
 * [3] Presentation for 08/03/07