A comparative study between the adsorption and covalent binding of human immunoglobulin and lysozyme on surface-modified poly(tert-butyl methacrylate).
The adsorption and covalent immobilization of human immunoglobulin (HIgG) and lysozyme (LYZ) on surface-modified poly(tert-butyl methacrylate) PtBMA films have been evaluated using x-ray photoelectron spectroscopy (XPS), ellipsometry and atomic force microscopy (AFM). Surface modification of PtBMA (UV irradiation) afforded surfaces suitable for both the physical and covalent attachment of proteins. The XPS and ellipsometry results showed good correlation in terms of variable-dense/thickness protein layer formation between physisorbed and covalently bound proteins. The amount of physisorbed HIgG ranged from 23.0 +/- 1.6 ng mm(2) on PtBMA, with corresponding film thicknesses 17.0 +/- 1.2 nm. Covalent immobilization mediated through 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysulfosuccinimide (sulfo-NHS) coupling chemistry, afforded 5.6-8 ng mm(2) of HIgG with a corresponding thickness of 5.9 +/- 0.6 nm on PtBMA. The attachment of LYZ to modified PtBMA surface was similarly translated, where adsorption yielded up to 15 ng mm(2), while covalent immobilization afforded typically 7-8 ng mm(2). The thickness of the adsorbed LYZ protein layer was 11.0 +/- 3.2 nm (PtBMA), suggesting the greater portion of protein adsorbs on surface-modified PtBMA.
Ivanova
Elena P
EP
Swinburne University of Technology, PO Box 218, Hawthorn, VIC 3122, Australia. eivanova@swin.edu.au
Wright
Jonathan P
JP
Pham
Duy K
DK
Brack
Narelle
N
Pigram
Paul
P
Alekseeva
Yulia V
YV
Demyashev
Gregory M
GM
Nicolau
Dan V
DV
D000042:Absorption; D001665:Binding Sites; D001672:Biocompatible Materials / Q000737:chemistry*; D004800:Enzymes, Immobilized; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry*; D008422:Materials Testing; D008689:Methacrylates / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry*; D011485:Protein Binding; D013499:Surface Properties
D001672:Biocompatible Materials; D004800:Enzymes, Immobilized; D007074:Immunoglobulin G; D008689:Methacrylates; D009113:Muramidase