反应详情

Cochleates containing OVA were prepared from unilamellar liposomes comprising a 9:1 mixture (w/w) of synthetic ester lipid 1,2-Dioleoyl-sn-Glycero-3-[Phospho-L-Serine] sodium salt (DOPS lyophilized powder; obtained from Avanti Polar Lipids Inc.; Alabaster, Ala., USA; MW of 810.04; cat #830035P, Lot #181PS-214) and cholesterol (Sigma Chemical Co.; Cat # C8667), by the standard cochleate method of direct addition of CaCl2 as described (Gould-Fogerite and Mannino, 1993). Briefly, a dried film of 20 mg mixture of the DOPS-cholesterol was hydrated in 1 ml of OVA stock solution (10 mg OVA/ml water) and converted to unilamellar liposomes by bath sonication, by avoiding exposure to air as described (Jin et al., 2000). Ten microliter aliquots of 0.1 M CaCl2 stock solution were added to the liposome suspension with mixing in between each addition, till the total concentration of calcium ions equaled to one half the molar concentration of DOPS constituting the liposomes, and then additional CaCl2 was added to achieve a 3 mM excess over that already added, as described (Gould-Fogerite and Mannino, 1993).

1. additionCochleates containing OVA
2. customwere prepared from unilamellar liposomes
3. customconverted to unilamellar liposomes by bath
4. customsonication
5. concentrationthe molar concentration of DOPS constituting the liposomes
6. additionadditional CaCl2 was added
7. additionthat already added