Effects of cloud-point grafting, chain length, and density of PEG layers on competitive adsorption of ocular proteins.
The effects of pinning density, chain length, and 'cloud point' (CP) versus non-CP grafting conditions have been studied on the ability of polyethylene glycol (PEG) layers to minimize adsorption from a multicomponent (lysozyme, human serum albumin (HSA), IgG and lactoferrin) protein solution. Methoxy-terminated aldehyde-PEG (M-PEG, MW 5000) and dialdehyde-PEG (PEG(ald)2, MW 3400) were grafted by reductive amination onto two surfaces of different amine group density, generated by radiofrequency glow discharge (r.f.g.d.) deposition of n-heptylamine (HA) (low density) or allylamine (AlA) (high density) r.f.g.d. polymer layers. The PEG graft density was varied also by increasing the temperature and salt (K2SO4) content of the grafting solution; it reached a maximum at the CP of the PEGs. The CP reaction conditions were critical for producing PEG layers capable of minimizing protein adsorption. X-ray photoelectron spectroscopy (XPS) showed that under these conditions, PEG(ald)2 produced a thick linear PEG layer, most likely by aldol condensation. Protein adsorption was assessed using XPS and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS) in the surface mode (Surface-MALDI-MS). Coatings grafted at non-CP conditions showed some protein adsorption, as did the HA/M-PEG layer grafted at the CP. On the other hand, no protein adsorption was detected on the HA/PEG(ald)2, AlA/M-PEG, and AlA/PEG(ald)2 surfaces when grafted at the CP. Thus, the effects of pinning density and chain length are interrelated, but the key factor is optimization of PEG chain density by use of the CP conditions, provided that a sufficient density of pinning sites exists.
Kingshott
Peter
P
CSIRO Molecular Science, Clayton, Vic, Australia. peter.kingshott@risoe.dk
Thissen
Helmut
H
Griesser
Hans J
HJ
D000327:Adsorption; D000499:Allylamine / Q000378:metabolism; D000588:Amines / Q000737:chemistry; D000818:Animals; D001667:Binding, Competitive; D002645:Chickens; D005123:Eye / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007477:Ions; D007781:Lactoferrin / Q000737:chemistry; D008422:Materials Testing; D009113:Muramidase / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013052:Spectrometry, X-Ray Emission; D013431:Sulfates / Q000494:pharmacology; D013499:Surface Properties; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature; D005719:gamma-Globulins / Q000737:chemistry
D000588:Amines; D007074:Immunoglobulin G; D007477:Ions; D011108:Polymers; D013431:Sulfates; D013501:Surface-Active Agents; D005719:gamma-Globulins; C031512:potassium sulfate; D011092:Polyethylene Glycols; D000499:Allylamine; D009113:Muramidase; D007781:Lactoferrin