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 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 0.1ng/ml have been measured with surface plasmon resonance.
Vikholm-Lundin
Inger
I
VTT Technical Research Centre of Finland, P.O. Box 1300, FI-33101 Tampere, Finland.
Pulli
Timo
T
Albers
Willem M
WM
Tappura
Kirsi
K
D000906:Antibodies / Q000235:genetics / Q000276:immunology*; D015374:Biosensing Techniques / Q000379:methods*; D007118:Immunoassay / Q000379:methods*; D007140:Immunoglobulin Fab Fragments / Q000032:analysis* / Q000276:immunology*; D015336:Molecular Probe Techniques; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D020349:Surface Plasmon Resonance / Q000379:methods*
D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins