Modification of liposomes with N-substituted polyacrylamides: identification of proteins adsorbed from plasma.
Liposomes prepared from DMPC (80%) and cholesterol (20%) were modified with a series of hydrophobically modified N-substituted polyacrylamides, namely, poly[N-isopropylacrylamide] (PNIPAM), poly[N,N-bis(2-methoxyethyl) acrylamide] (PMEAM), and poly[(3-methoxypropyl)acrylamide] (PMPAM). The hydrophobic group, N-[4-(1-pyrenylbutyl)-N-n-octadecylamine was attached to one end of the polymer chains to serve as an anchor for incorporation into the liposome bilayer. Liposome-polymer interactions were confirmed using fluorescence spectroscopy and chemical analysis. Microscopy revealed differences in aggregation tendency between unmodified and polymer-modified liposomes. Proteins adsorbed to liposome surfaces during exposure to human plasma were identified by immunoblot analysis. It was found that both unmodified and polymer-modified liposomes adsorb a wide variety of plasma proteins. Contact phase coagulation proteins, complement proteins, cell-adhesive proteins, serine protease inhibitors, plasminogen, antithrombin III, prothrombin, transferrin, alpha(2)-microglobulin, hemoglobin, haptoglobin and beta-lipoprotein as well as the major plasma proteins were all detected. Some differences were found between the unmodified and polymer-modified liposomes. The unmodified liposomes adsorbed plasminogen mainly as the intact protein, whereas on the modified liposomes plasminogen was present in degraded form. Also, the liposomes modified with PNIPAM in its extended conformation (below the lower critical solution temperature) appeared to adsorb less protein than those containing the 'collapsed' form of PNIPAM (above the LCST).
Yamazaki
A
A
Department of Chemical Engineering, McMaster University, Hamilton, ON, Canada.
Winnik
F M
FM
Cornelius
R M
RM
Brash
J L
JL
D000178:Acrylamides / Q000737:chemistry*; D000327:Adsorption; D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry*; D015153:Blotting, Western; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D008968:Molecular Conformation; D015394:Molecular Structure; D011108:Polymers / Q000737:chemistry*; D013696:Temperature; D014867:Water / Q000737:chemistry
D000178:Acrylamides; D001798:Blood Proteins; D004337:Drug Carriers; D008081:Liposomes; D011108:Polymers; D014867:Water; D004134:Dimyristoylphosphatidylcholine