title: Functional characterisation of Fab'-fragments self-assembled onto hydrophilic gold surfaces.
abstract: Antibody Fab'-fragments have been immobilised on hydrophilic gold by direct self-assembly, and embedded in a matrix of non-ionic hydrophilic polymers, tris(hydroxymethyl)methylacrylamide, carrying lipoate terminal linking groups. Different polymers were synthesised, and co-adsorbed or post-adsorbed between the antibody fragments in order to optimise the antigen binding. Various factors were investigated that influence the activity of the immobilised Fab'-fragments for binding of the antigen, human IgG. The Fab'-fragments were immobilised in dense layers close to monolayer coverage, and the stoichiometric efficiency of immobilisation was up to 30%, with the human IgG also approaching monolayer coverage. The cleaning of the gold surface was a crucial factor in preservation of activity. Besides the usual treatment in hot ammonia/peroxide solution, hot DMSO appeared to be highly effective as a cleaning agent.
authors.affiliation: VTT Technical Research Centre of Finland, P.O. Box 1300, FI-33101 Tampere, Finland. martin.albers@vtt.fi
authors.forename: Willem M
authors.initials: WM
authors.lastname: Albers
authors.forename: Sanna
authors.initials: S
authors.lastname: Auer
authors.forename: Hannu
authors.initials: H
authors.lastname: Helle
authors.forename: Tony
authors.initials: T
authors.lastname: Munter
authors.forename: Inger
authors.initials: I
authors.lastname: Vikholm-Lundin
mesh_terms: D000327:Adsorption; D006046:Gold / Q000737:chemistry*; D006801:Humans; D055767:Immobilized Proteins / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D007700:Kinetics; D011108:Polymers / Q000737:chemistry; D012015:Reference Standards; D020349:Surface Plasmon Resonance; D013499:Surface Properties
chemical_list: D055767:Immobilized Proteins; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D011108:Polymers; D006046:Gold