Stabilized plasmid-lipid particles: construction and characterization.
A detergent dialysis procedure is described which allows encapsulation of plasmid DNA within a lipid envelope, where the resulting particle is stabilized in aqueous media by the presence of a poly(ethyleneglycol) (PEG) coating. These 'stabilized plasmid-lipid particles' (SPLP) exhibit an average size of 70 nm in diameter, contain one plasmid per particle and fully protect the encapsulated plasmid from digestion by serum nucleases and E. coli DNase I. Encapsulation is a sensitive function of cationic lipid content, with maximum entrapment observed at dioleoyldimethylammonium chloride (DODAC) contents of 5 to 10 mol%. The formulation process results in plasmid-trapping efficiencies of up to 70% and permits inclusion of 'fusigenic' lipids such as dioleoylphosphatidylethanolamine (DOPE). The in vitro transfection capabilities of SPLP are demonstrated to be strongly dependent on the length of the acyl chain contained in the ceramide group used to anchor the PEG polymer to the surface of the SPLP. Shorter acyl chain lengths result in a PEG coating which can dissociate from the SPLP surface, transforming the SPLP from a stable particle to a transfection-competent entity. It is suggested that SPLP may have utility as systemic gene delivery systems for gene therapy protocols.
Wheeler
J J
JJ
Inex Pharmaceuticals Corporation, Burnaby, BC, Canada.
Palmer
L
L
Ossanlou
M
M
MacLachlan
I
I
Graham
R W
RW
Zhang
Y P
YP
Hope
M J
MJ
Scherrer
P
P
Cullis
P R
PR
D000818:Animals; D019556:COS Cells; D002214:Capsules; D002460:Cell Line; D005614:Freeze Fracturing; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008081:Liposomes; D008854:Microscopy, Electron; D010957:Plasmids; D011092:Polyethylene Glycols; D014162:Transfection / Q000379:methods*
D002214:Capsules; D008081:Liposomes; D011092:Polyethylene Glycols