TY - JOUR
T1 - Surface immobiliastion of antibody on cyclic olefin copolymer for sandwich immunoassay
AU - RAJ, Jog
AU - HERZOG, Grégoire
AU - MANNING, Mary
AU - Volcke, Cedric
AU - MacCRAITH, Brian D.
AU - BALLANTYNE, Scott
AU - THOMPSON, Michael
AU - ARRIGAN, Damien W.M.
PY - 2009
Y1 - 2009
N2 - There is a growing interest in the use of plastic materials in the development of biochips for various applications. In this work, the surface functionalisation of the commercially-available cyclic olefin co-polymer materials Zeonor (ZR) and Zeonex (ZX) was examined. The methodology developed entailed oxidation in oxygen plasma, functionalisation of the oxidized surface with aminopropyl triethoxy silane and attachment of antibodies using covalent linker molecules, namely 1,4-Phenylene diisothiocyanate (PDITC), Toluene-2,4-Diisocyanate and Sulfo-Succinimidyl-4-(N-Maleimidomethyl) cyclohexane-1-carboxylate. PDITC was selected as the most suitable cross-linker for the attachment of antibodies, as assessed by fluorescent intensity measurements on immobilised FITC-labelled IgG antibodies. The modification method was characterised by contact angle measurements, ellipsometry, x-ray photoelectron spectroscopy (XPS) and fluorescence microscopy. The data are consistent with the deposition of a polymeric film of APTES chemisorbed to the oxidised plastic surface. The functionalised surfaces were employed in a sandwich immunoassay format using the reagents goat anti-human IgG (GantiHIgG) and fluorescently-labelled GantiHIgG (Cy5-GantiHIgG) as capture and detection antibodies, respectively, and with human IgG (HIgG) as the model analyte. The lowest concentration of HIgG detected was 0.2 ng ml-1, with a relative standard deviation of 15%. Non-specific binding effects were also assessed. The method and supporting data demonstrate that simple approaches to surface functionalisation can be adapted to plastic chips.
AB - There is a growing interest in the use of plastic materials in the development of biochips for various applications. In this work, the surface functionalisation of the commercially-available cyclic olefin co-polymer materials Zeonor (ZR) and Zeonex (ZX) was examined. The methodology developed entailed oxidation in oxygen plasma, functionalisation of the oxidized surface with aminopropyl triethoxy silane and attachment of antibodies using covalent linker molecules, namely 1,4-Phenylene diisothiocyanate (PDITC), Toluene-2,4-Diisocyanate and Sulfo-Succinimidyl-4-(N-Maleimidomethyl) cyclohexane-1-carboxylate. PDITC was selected as the most suitable cross-linker for the attachment of antibodies, as assessed by fluorescent intensity measurements on immobilised FITC-labelled IgG antibodies. The modification method was characterised by contact angle measurements, ellipsometry, x-ray photoelectron spectroscopy (XPS) and fluorescence microscopy. The data are consistent with the deposition of a polymeric film of APTES chemisorbed to the oxidised plastic surface. The functionalised surfaces were employed in a sandwich immunoassay format using the reagents goat anti-human IgG (GantiHIgG) and fluorescently-labelled GantiHIgG (Cy5-GantiHIgG) as capture and detection antibodies, respectively, and with human IgG (HIgG) as the model analyte. The lowest concentration of HIgG detected was 0.2 ng ml-1, with a relative standard deviation of 15%. Non-specific binding effects were also assessed. The method and supporting data demonstrate that simple approaches to surface functionalisation can be adapted to plastic chips.
KW - APTES
KW - Cyclic olefin copolymers
KW - surface functionalisation
KW - antibody immobilization
KW - immunoassay
M3 - Article
SN - 0956-5663
VL - 24
SP - 2654
EP - 2658
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
ER -