Highly efficient immobilisation of antibody fragments to functionalised lipid monolayers

Inger Vikholm (Corresponding Author), Tapani Viitala, Martin Albers, Jouko Peltonen

Research output: Contribution to journalArticleScientificpeer-review

37 Citations (Scopus)

Abstract

The covalent attachment of Fab′ fragments of polyclonal anti-human IgG to a lipid with a terminal linker group was examined by means of quartz crystal microbalance and surface plasmon resonance measurements. The linker lipid was embedded in binary or ternary monolayers of dipalmitoylphosphatidylcholine (DPPC) and cholesterol. Atomic force microscopy images of the films deposited on silanised SiO2 substrates showed that Fab′ fragments take a standing position, thus giving site-directed immobilisation. Human IgG forms a network on interaction with the antibodies. Non-specific binding of bovine serum albumin was found to be very low when DPPC was used as the host matrix. At an optimal Fab′ fragment concentration a binding capacity above 60% was obtained. However, if the surface concentration of the immobilised antibodies was too high, the binding capacity decreased due to steric hindrance. The results demonstrate that the covalent coupling of Fab′ fragments to N-(ϵ-maleimidocaproyl)-dipalmitoylphosphatidylethanolamine (DPPE-EMC) embedded in a host monolayer matrix of DPPC is a promising approach to achieve a site-directed immobilisation of antibodies with high antigen-binding efficiency.

Original languageEnglish
Pages (from-to)39-52
Number of pages14
JournalBiochimica et Biophysica Acta
Volume1421
Issue number1
DOIs
Publication statusPublished - 1999
MoE publication typeA1 Journal article-refereed

Fingerprint

Immunoglobulin Fragments
Immunoglobulin Fab Fragments
1,2-Dipalmitoylphosphatidylcholine
Immobilization
Monolayers
Lipids
Immunoglobulin G
Quartz Crystal Microbalance Techniques
Immobilized Antibodies
Surface Plasmon Resonance
Antibodies
Atomic Force Microscopy
Quartz crystal microbalances
Electromagnetic compatibility
Surface plasmon resonance
Bovine Serum Albumin
Posture
Atomic force microscopy
Cholesterol
Antigens

Cite this

Vikholm, Inger ; Viitala, Tapani ; Albers, Martin ; Peltonen, Jouko. / Highly efficient immobilisation of antibody fragments to functionalised lipid monolayers. In: Biochimica et Biophysica Acta. 1999 ; Vol. 1421, No. 1. pp. 39-52.
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abstract = "The covalent attachment of Fab′ fragments of polyclonal anti-human IgG to a lipid with a terminal linker group was examined by means of quartz crystal microbalance and surface plasmon resonance measurements. The linker lipid was embedded in binary or ternary monolayers of dipalmitoylphosphatidylcholine (DPPC) and cholesterol. Atomic force microscopy images of the films deposited on silanised SiO2 substrates showed that Fab′ fragments take a standing position, thus giving site-directed immobilisation. Human IgG forms a network on interaction with the antibodies. Non-specific binding of bovine serum albumin was found to be very low when DPPC was used as the host matrix. At an optimal Fab′ fragment concentration a binding capacity above 60{\%} was obtained. However, if the surface concentration of the immobilised antibodies was too high, the binding capacity decreased due to steric hindrance. The results demonstrate that the covalent coupling of Fab′ fragments to N-(ϵ-maleimidocaproyl)-dipalmitoylphosphatidylethanolamine (DPPE-EMC) embedded in a host monolayer matrix of DPPC is a promising approach to achieve a site-directed immobilisation of antibodies with high antigen-binding efficiency.",
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Highly efficient immobilisation of antibody fragments to functionalised lipid monolayers. / Vikholm, Inger (Corresponding Author); Viitala, Tapani; Albers, Martin; Peltonen, Jouko.

In: Biochimica et Biophysica Acta, Vol. 1421, No. 1, 1999, p. 39-52.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Highly efficient immobilisation of antibody fragments to functionalised lipid monolayers

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AU - Viitala, Tapani

AU - Albers, Martin

AU - Peltonen, Jouko

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PY - 1999

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AB - The covalent attachment of Fab′ fragments of polyclonal anti-human IgG to a lipid with a terminal linker group was examined by means of quartz crystal microbalance and surface plasmon resonance measurements. The linker lipid was embedded in binary or ternary monolayers of dipalmitoylphosphatidylcholine (DPPC) and cholesterol. Atomic force microscopy images of the films deposited on silanised SiO2 substrates showed that Fab′ fragments take a standing position, thus giving site-directed immobilisation. Human IgG forms a network on interaction with the antibodies. Non-specific binding of bovine serum albumin was found to be very low when DPPC was used as the host matrix. At an optimal Fab′ fragment concentration a binding capacity above 60% was obtained. However, if the surface concentration of the immobilised antibodies was too high, the binding capacity decreased due to steric hindrance. The results demonstrate that the covalent coupling of Fab′ fragments to N-(ϵ-maleimidocaproyl)-dipalmitoylphosphatidylethanolamine (DPPE-EMC) embedded in a host monolayer matrix of DPPC is a promising approach to achieve a site-directed immobilisation of antibodies with high antigen-binding efficiency.

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