Tak 242 cli 095

1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) lipid monomers were purchased from Avanti Polar Lipids, Inc. TAK-242 (CLI-095) was purchased from Invivogen. Lipids with or without TAK-242 were dissolved in chloroform and mixed. The chloroform was evaporated under nitrogen, and the lipid film was subsequently lyophilized. The lipid film was then rehydrated in a buffer containing 20 mM Tris-HCL and 150 mM NaCl. The lipid solution was probe sonicated at an amplitude of 40% for 30−40 min (QSonica). Following sonication, lipids were centrifuged at 4000 rpm for 90 min to remove larger lipid vesicles. The supernatant was removed and extruded through polycarbonate membranes of increasingly smaller pore size (80, 50, and 30 nm), until the resulting lipid particles were monodisperse, ascertained by dynamic light scattering (Malvern). The particles were passed through a column containing cross-linked Sepharose to remove any unencapsulated material. The concentration of lipids and encapsulated TAK-242 were determined by inductively coupled plasma optical emission spectroscopy (Thermo iCAP 7600). The 1.3 mM lipids were mixed with 15 μM cholesterol-terminated DNA for 3−4 h at 37 °C to allow for cholesterol intercalation into the lipid bilayer of the liposomes. Size and polydispersity were measured by DLS.