A comparative evaluation of molecular recognition by monolayers composed of synthetic receptors or oriented antibodies

Recombinant anti-morphine Fab′ fragments have been immobilised on gold by covalent attachment through the free thiol groups of the fragment. The antibody fragments were intercalated with a non-ionic hydrophilic polymer in order to suppress non-specific binding of interfering substances. The antibodi...

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Veröffentlicht in:Biosensors & bioelectronics 2008-12, Vol.24 (4), p.1036-1038
Hauptverfasser: Vikholm-Lundin, Inger, Pulli, Timo, Albers, Willem M., Tappura, Kirsi
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container_issue 4
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container_title Biosensors & bioelectronics
container_volume 24
creator Vikholm-Lundin, Inger
Pulli, Timo
Albers, Willem M.
Tappura, Kirsi
description Recombinant anti-morphine Fab′ fragments have been immobilised on gold by covalent attachment through the free thiol groups of the fragment. The antibody fragments were intercalated with a non-ionic hydrophilic polymer in order to suppress non-specific binding of interfering substances. The antibodies are oriented on the surface due to the thiol groups of the antibody and the layer shows a high response to antigen. Non-specific binding of bovine serum albumin is moreover very low because of the repellent polymer. Synthetic receptors composed of an imprinted self-assembled monolayer made from lipoates and the template, morphine, exhibit the same binding response to the antigen, morphine as the site-specific oriented antibody monolayer. A similar binding curve could be obtained as that for binding of morphine to an antibody Fab′ fragment/polymer layer – indicating that synthetic receptors produced are comparable to those of antibody layers. Concentrations down to 0.1 ng/ml have been measured with surface plasmon resonance.
doi_str_mv 10.1016/j.bios.2008.06.049
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subjects Antibodies - genetics
Antibodies - immunology
Antibody Fab′ fragment
Biosensing Techniques - methods
Immobilisation
Immunoassay
Immunoassay - methods
Immunoglobulin Fab Fragments - analysis
Immunoglobulin Fab Fragments - immunology
Imprinted self-assembled monolayer
Molecular Probe Techniques
Protein repellent polymer
Recombinant Proteins - analysis
Recombinant Proteins - immunology
Surface plasmon resonance
Surface Plasmon Resonance - methods
title A comparative evaluation of molecular recognition by monolayers composed of synthetic receptors or oriented antibodies
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