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

Willem M. Albers (Corresponding Author), Sanna Auer, Hannu Helle, Tony Munter, Inger Vikholm-Lundin

Research output: Contribution to journalArticleScientificpeer-review

10 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 languageEnglish
Pages (from-to)193-199
JournalColloids and Surfaces B: Biointerfaces
Volume68
Issue number2
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fingerprint

Immunoglobulin Fab Fragments
Antigens
Antibodies
Gold
Cleaning
Monolayers
Immunoglobulin Fragments
fragments
gold
Polymers
antigens
Peroxides
Immunoglobulin G
antibodies
Self assembly
cleaning
Ammonia
Dimethyl Sulfoxide
Immobilization
polymers

Keywords

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

Cite this

Albers, Willem M. ; Auer, Sanna ; Helle, Hannu ; Munter, Tony ; Vikholm-Lundin, Inger. / Functional characterisation of Fab'-fragments self-assembled onto hydrophilic gold surfaces. In: Colloids and Surfaces B: Biointerfaces. 2009 ; Vol. 68, No. 2. pp. 193-199.
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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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Functional characterisation of Fab'-fragments self-assembled onto hydrophilic gold surfaces. / Albers, Willem M. (Corresponding Author); Auer, Sanna; Helle, Hannu; Munter, Tony; Vikholm-Lundin, Inger.

In: Colloids and Surfaces B: Biointerfaces, Vol. 68, No. 2, 2009, p. 193-199.

Research output: Contribution to journalArticleScientificpeer-review

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

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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

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JO - Colloids and Surfaces B: Biointerfaces

JF - Colloids and Surfaces B: Biointerfaces

SN - 0927-7765

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