Eva Österberg

Comparison of polysaccharide and poly(ethylene glycol) coatings for reduction of protein adsorption on polystyrene surfaces

Abstract There has been much recent interest in the use of poly(ethylene glycol)s (PEGs) for a variety of biotechnical applications. In the present work we have immobilized several cellulose derivatives and dextran on polystyrene surfaces and have measured the extent of fibrinogen adsorption onto the coated surfaces. Immobilization was achieved by adsorption onto clean polystyrene and by covalent linkage of oxidized polysaccharides to polyethylenimine which was ionically bound to polystyrene. Covalently bound polysaccharides, and adsorbed polysaccharides that are strongly held, compare well with poly(ethylene glycol) in preventing fibrinogen adsorption. The same polymers were coupled to polystyrene latex particles to permit examination by analytical microparticle electrophoresis. This investigation suggests that adsorbed polysaccharides form thicker layers than do covalently bound polysaccharides. Despite the polysaccharides being bound at many points along the polymer chain while PEG is bound only at the polymer terminus, the functional equivalence of polysaccharide and PEG coatings is of significance in interpreting the protein-rejecting ability of polymer-modified surfaces.

Effects of branching and molecular weight of surface-bound poly(ethylene oxide) on protein rejection.

To understand better the origin of protein rejection observed with surface-bound poly(ethylene oxide) (or PEO), we have measured fibrinogen adsorption for a series of linear and branched, low-molecular-weight PEOs bound to solid polystyrene surfaces. The results show that a dependence on molecular weight is found below 1500 g mol-1 for linear PEO. Branched PEOs are less effective at protein rejection than linear PEOs. The branched PEOs have smaller exclusion volumes (from GPC) than the corresponding linear PEOs, consistent with restriction in conformational freedom for the branched compounds. The protein rejection results are interpreted in terms of entropy changes that result upon protein adsorption. In addition, some practical problems in preparation of PEO glycidyl ethers have been clarified, thus making these PEO derivatives more useful for surface modification.

Synthesis of phosphatidylcholine with (n-3) fatty acids by phospholipase A 2 in microemulsion

Phosphatidylcholine containing a long chain polyunsaturated acyl group at the 2-position has been prepared by phospholipase A 2 catalyzed esterification of lysophosphatidylcholine with polyunsaturated fatty acids EPA (C20∶5) or DHA (C22∶6). Preliminary studies showed that the other fatty acids, such as lauric acid (C12), palmitic acid (C16), stearic acid (C18) and linoleic acid (C18∶2), were also incorporated. To our knowledge, phospholipase A 2 catalyzed condensation reactions have not been reported in the literature before. The reactions were performed in sodiumbis(2-ethylhexyl)-sulfosuccinate-based microemulsions containing small amounts of water. Synthesis of the same phospholipid by transesterification of phosphatidylcholine with the polyunsaturated acids in microemulsion failed; however, enzymatic hydrolysis to lysophosphatidylcholine was facile, quantitative conversion from phosphatidylcholine being attained after 16 hr reaction time.An additional observation was that, unlike enzymatic hydrolysis of phospholipids, the condensation reaction catalyzed by phospholipase A 2 was totally independent of the presence of calcium.

Using poly(ethylene imine) to graft poly(ethylene glycol) or polysaccharide to polystyrene

Abstract A novel method of grafting poly(ethylene glycol) (PEG) or polysaccharide to polystyrene has been developed. The non-charged, hydrophilic polymer is firstly grafted to poly(ethylene imine). After prior oxidation of the solid surface, the graft copolymer is subsequently adsorbed. Analysis by electron spectroscopy for chemical analysis and ellipsometry indicates a trains-and-loops arrangement of the copolymer on the surface with densely packed PEG or polysaccharide chains oriented towards the bulk water phase. Ellipsometry also shows that whereas the PEG grafting is fast, the polysaccharide grafting is more sluggish. Attachment of both graft copolymers is completely irreversible, as seen also by ellipsometry. The surfaces obtained are highly protein repellent, as shown by enzyme-linked immunosorbent assay using fibrinogen and immunoglobulin G as model proteins.