Functional characterisation of Fab'-fragments self-assembled onto hydrophilic gold surfaces

Willem M. Albers; Sanna Auer; Hannu Helle; Tony Munter; Inger Vikholm-Lundin

doi:10.1016/j.colsurfb.2008.10.001

Functional characterisation of Fab'-fragments self-assembled onto hydrophilic gold surfaces

Willem M. Albers^*
, Sanna Auer
, Hannu Helle
, Tony Munter
, Inger Vikholm-Lundin

^*Corresponding author for this work

VTT Technical Research Centre of Finland

VTT (former employee or external)

Research output: Contribution to journal › Article › Scientific › peer-review

16 Citations (Scopus)

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.

Original language	English
Pages (from-to)	193-199
Journal	Colloids and Surfaces B: Biointerfaces
Volume	68
Issue number	2
DOIs	https://doi.org/10.1016/j.colsurfb.2008.10.001
Publication status	Published - 2009
MoE publication type	A1 Journal article-refereed

Keywords

Immunoassay
Heterogeneous binding
Surface plasmon resonance
Antibody Fab'-fragment
Immobilisation

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10.1016/j.colsurfb.2008.10.001

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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.",

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author = "Albers, \{Willem M.\} and Sanna Auer and Hannu Helle and Tony Munter and Inger Vikholm-Lundin",

year = "2009",

doi = "10.1016/j.colsurfb.2008.10.001",

language = "English",

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issn = "0927-7765",

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TY - JOUR

T1 - Functional characterisation of Fab'-fragments self-assembled onto hydrophilic gold surfaces

AU - Albers, Willem M.

AU - Auer, Sanna

AU - Helle, Hannu

AU - Munter, Tony

AU - Vikholm-Lundin, Inger

PY - 2009

Y1 - 2009

N2 - 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.

AB - 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.

KW - Immunoassay

KW - Heterogeneous binding

KW - Surface plasmon resonance

KW - Antibody Fab'-fragment

KW - Immobilisation

U2 - 10.1016/j.colsurfb.2008.10.001

DO - 10.1016/j.colsurfb.2008.10.001

M3 - Article

SN - 0927-7765

VL - 68

SP - 193

EP - 199

JO - Colloids and Surfaces B: Biointerfaces

JF - Colloids and Surfaces B: Biointerfaces

IS - 2

ER -

Functional characterisation of Fab'-fragments self-assembled onto hydrophilic gold surfaces

Abstract

Keywords

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