Protein adsorption to polyethylene glycol modified liposomes from fibrinogen solution and from plasma.
Unmodified and polyethylene glycol (PEG) modified neutral and negatively charged liposomes were prepared by freeze-thaw and extrusion followed by chromatographic purification. The effects of PEG molecular weight (PEG 550, 2000, 5000), PEG loading (0-15 mol%), and liposome surface charge on fibrinogen adsorption were quantified using radiolabeling techniques. All adsorption isotherms increased monotonically over the concentration range 0-3 mg/ml and adsorption levels were low. Negatively charged liposomes adsorbed significantly more fibrinogen than neutral liposomes. PEG modification had no effect on fibrinogen adsorption to neutral liposomes. An inverse relationship was found between PEG loading of negatively charged liposomes and fibrinogen adsorption. PEGs of all three molecular weights at a loading of 5 mol% reduced fibrinogen adsorption to negatively charged liposomes. Protein adsorption from diluted plasma (10% normal strength) to four different liposome types (neutral, PEG-neutral, negatively charged, and PEG-negatively charged) was investigated using gel electrophoresis and immunoblotting. The profiles of adsorbed proteins were similar on all four liposome types, but distinctly different from the profile of plasma itself, indicating a partitioning effect of the lipid surfaces. alpha2-macroglobulin and fibronectin were significantly enriched on the liposomes whereas albumin, transferrin, and fibrinogen were depleted compared to plasma. Apolipoprotein AI was a major component of the adsorbed protein layers. The blot of complement protein C3 adsorbed on the liposomes suggested that the complement system was activated.
Price
M E
ME
Department of Chemical Engineering, McMaster University, 1280 Main Street West, L8S 4L7, Hamilton, ON, Canada.
Cornelius
R M
RM
Brash
J L
JL
D000327:Adsorption; D001779:Blood Coagulation Factors / Q000737:chemistry / Q000302:isolation & purification; D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification; D015153:Blotting, Western; D002784:Cholesterol / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry*; D008970:Molecular Weight; D010713:Phosphatidylcholines / Q000737:chemistry*; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D010715:Phosphatidylglycerols / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013499:Surface Properties
D001779:Blood Coagulation Factors; D001798:Blood Proteins; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D011092:Polyethylene Glycols; D005340:Fibrinogen; D002784:Cholesterol; C002773:dimyristoylphosphatidylglycerol