Irreversible adsorption of human serum albumin to hydrogel contact lenses: a study using electron spin resonance spectroscopy.
Human serum albumin (HSA) was specifically spin labelled with 4-maleimido-tempo (MSL) at its cysteine 34 residue (HSA-MSL). The irreversible adsorption of HSA-MSL to hydrogel contact lenses (etafilcon A, tefilcon and vifilcon A) was investigated using electron spin resonance (ESR) spectroscopy. Changes in ESR spectral characteristics of adsorbed HSA-MSL as compared to HSA-MSL in solution displayed an additional immobilisation of the spin label due to the adsorption. This immobilisation of MSL corresponds to a large conformational alteration of the HSA-MSL near the modified Cys 34 residue. For both etafilcon A and tefilcon, the rate of irreversible adsorption was relatively slow compared with that of vifilcon A where the maximum state of immobilisation and hence conformational change occurred within the first hour of adsorption. Furthermore, tefilcon produced markedly different ESR spectra where a strong conformational change to a less mobile protein was apparent. This supported a model where the direct irreversible adsorption of HSA from solution dominated on tefilcon as opposed to conversion of the adsorbed protein from the reversible to the irreversible state on both etafilcon A and vifilcon A. HSA-MSL adsorption onto hydrophobic poly(methylmethacrylate) (PMMA) and hydrophilic poly(N-ter-butylacrylamide) (PTBAM) latex beads was also investigated. The spin label MSL was found to be less mobile when HSA was adsorbed onto PMMA compared with PTBAM beads. It was also found that the rate of irreversible adsorption of HSA is far higher onto PMMA surfaces than onto PTBAM surfaces.
Cooperative Research Centre for Eye Research and Technology, University of New South Wales, Sydney, Australia. q.garrett@cclru.unsw.edu.au
Q
Q
Garrett
H J
HJ
Griesser
B K
BK
Milthorpe
R W
RW
Garrett
D000327:Adsorption; D003263:Contact Lenses, Hydrophilic; D003497:Cyclic N-Oxides / Q000737:chemistry*; D003545:Cysteine; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D007700:Kinetics; D011487:Protein Conformation; D012709:Serum Albumin / Q000737:chemistry*; D013113:Spin Labels; D013997:Time Factors
D003497:Cyclic N-Oxides; D020100:Hydrogels; D012709:Serum Albumin; D013113:Spin Labels; C017480:4-maleimido-2,2,6,6-tetramethylpiperidinooxyl; D003545:Cysteine