Biofunctionalized protein resistant oligo(ethylene glycol)-derived polymer brushes as selective immobilization and sensing platforms.
Poly(oligo(ethylene glycol) methacrylate) (POEGMA) brushes are extremely protein resistant polymer coatings that can reduce nonspecific adsorption of proteins from complex mixtures such as blood, sera and plasma. These coatings can be prepared via atom transfer radical polymerization with excellent control of their thickness and grafting density. We studied their direct functionalization with streptavidin and developed an assay for determining which coupling conditions afford the highest streptavidin loading efficiency. Disuccinimidyl carbonate was found to be the most efficient activating agent for covalent capture of the receptor. Using infrared and X-ray photoelectron spectroscopy, fluorescence microscopy, surface plasmon resonance, and ellipsometry, we examined how structural parameters such as the length of the oligo(ethylene glycol) side chain affect streptavidin functionalization, but also immobilization of biotinylated antibodies, subsequent selective secondary recognition and nonspecific binding of proteins. We found evidence that large macromolecules cannot infiltrate dense polymer brushes and that bulky antibody recognition occurs in the upper part of these coatings.
Trmcic-Cvitas
Jelena
J
Department of Chemistry, University of Cambridge, Cambridge, United Kingdom.
Hasan
Erol
E
Ramstedt
Madeleine
M
Li
Xin
X
Cooper
Matthew A
MA
Abell
Chris
C
Huck
Wilhelm T S
WT
Gautrot
Julien E
JE
D019855:Ethylene Glycol / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D013057:Spectrum Analysis; D020349:Surface Plasmon Resonance; D014965:X-Rays
D011108:Polymers; D011506:Proteins; D019855:Ethylene Glycol