Force probe measurements of antibody-antigen interactions

Deborah E Leckband, T. L. Kuhl, H. K. Wang, W. Müller, J. Herron, H. Ringsdorf

Research output: Contribution to journalArticle

Abstract

The surface force apparatus has been used to quantify directly the forces that govern the interactions between proteins and ligands. In this work, we describe the measured interactions between the antigen fluorescein and the Fab′ fragment of the monoclonal 4-4-20 anti-fluorescyl IgG antibody. Here we first describe the use of the surface force apparatus to demonstrate directly the impact of the charge composition in the region of the antibody binding site on the antibody interactions. Several approaches are described for immobilizing antigens, antibodies, and proteins in general for direct force measurements. The measured force profiles presented are accompanied by an extensive discussion of protocols used to analyze the force - distance curves and to interpret them in terms of the antibody structure. In addition to long-range electrostatic forces, we also consider short-range forces that can affect the strength of adhesion between the Fab′ and immobilized fluorescein. The latter investigations demonstrate the influence of interfacial properties on the recognition of surface-bound antigens.

Original languageEnglish (US)
Pages (from-to)329-340
Number of pages12
JournalMethods
Volume20
Issue number3
DOIs
StatePublished - Jan 1 2000

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Antigens
Antibodies
Fluorescein
Antibody Binding Sites
Immunoglobulin Fab Fragments
Electrostatic force
Force measurement
Surface Antigens
Static Electricity
Proteins
Adhesion
Ligands
Chemical analysis
anti-IgG

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Leckband, D. E., Kuhl, T. L., Wang, H. K., Müller, W., Herron, J., & Ringsdorf, H. (2000). Force probe measurements of antibody-antigen interactions. Methods, 20(3), 329-340. https://doi.org/10.1006/meth.1999.0926

Force probe measurements of antibody-antigen interactions. / Leckband, Deborah E; Kuhl, T. L.; Wang, H. K.; Müller, W.; Herron, J.; Ringsdorf, H.

In: Methods, Vol. 20, No. 3, 01.01.2000, p. 329-340.

Research output: Contribution to journalArticle

Leckband, DE, Kuhl, TL, Wang, HK, Müller, W, Herron, J & Ringsdorf, H 2000, 'Force probe measurements of antibody-antigen interactions', Methods, vol. 20, no. 3, pp. 329-340. https://doi.org/10.1006/meth.1999.0926
Leckband DE, Kuhl TL, Wang HK, Müller W, Herron J, Ringsdorf H. Force probe measurements of antibody-antigen interactions. Methods. 2000 Jan 1;20(3):329-340. https://doi.org/10.1006/meth.1999.0926
Leckband, Deborah E ; Kuhl, T. L. ; Wang, H. K. ; Müller, W. ; Herron, J. ; Ringsdorf, H. / Force probe measurements of antibody-antigen interactions. In: Methods. 2000 ; Vol. 20, No. 3. pp. 329-340.
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