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Quantitative interpretation of gold nanoparticle-based bioassays designed for detection of immunocomplex formation

Author:
  • Ye Zhou
  • Hongxing Xu
  • Andreas Dahlin
  • Jacob Vallkil
  • Carl Borrebaeck
  • Christer Wingren
  • Bo Liedberg
  • Fredrik Höök
Publishing year: 2007
Language: English
Pages: 6-15
Publication/Series: Biointerphases
Volume: 2
Issue: 1
Document type: Journal article
Publisher: AVS

Abstract english

The authors present in this paper how the extended Mie theory can be used to translate not only end-point data but also temporal variations of extinction peak-position changes, peak(t), into absolute mass uptake, (t), upon biomacromolecule binding to localized surface plasmon resonance (SPR) active nanoparticles (NPs). The theoretical analysis is applied on a novel sensor template composed of a three-layer surface architecture based on (i) a self-assembled monolayer of HS(CH2)15COOH, (ii) a 1:1 mixture of biotinylated and pure poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG), and (iii) NeutrAvidin. Assisted by independent estimations of the thickness of the three-layer architecture using quartz crystal microbalance with dissipation (QCM-D) monitoring, excellent agreement with parallel mass-uptake estimations using planar SPR is obtained. Furthermore, unspecific binding of serum to PLL-g-PEG was shown to be below the detection limit, making the surface architecture ideally suited for label-free detection of immunoreactions. To ensure that the immunocomplex formation occurred within the limited sensing depth (~10 nm) of the NPs, a compact model system composed of a biotinylated human recombinant single-chain antibody fragment (~2 nm) directed against cholera toxin was selected. By tracking changes in the centroid (center of mass) of the extinction peak, rather than the actual peak position, signal-to-noise levels and long-term stability upon cholera toxin detection are demonstrated to be competitive with results obtained using conventional SPR and state-of-the-art QCM-D data.

Keywords

  • Condensed Matter Physics
  • Immunology in the medical area

Other

Published
  • ISSN: 1934-8630
Carl B
Carl Borrebaeck
E-mail: carl [dot] borrebaeck [at] immun [dot] lth [dot] se

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Department of Immunotechnology

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