Characterization of DNA Immobilization and Subsequent Hybridization on a 2D Arrangement of Streptavidin on a Biotin-Modified Lipid Bilayer Supported on SiO₂

Abstract

We show how the water content (and effective density) of thin adsorbed films composed of biomolecules can be determined using combined quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) analysis. In particular, these techniques, combined with theoretical treatment using a Voigt-based viscoelastic model, were applied to analyze the state of surface immobilized single stranded biotin-modified probe DNA (b-DNA) coupled via streptavidin to a biotin-doped supported phospholipid bilayer (b-SPB)). From a proper analysis, it is demonstrated how changes in effective thickness, delta(f), and the viscoelastic components (shear viscosity, eta(f), and shear elasticity, mu(f))) can be obtained during both DNA immobilization and hybridization with single stranded fully complementary target DNA. In particular, it is demonstrated how this type of analysis can be used to control the state of streptavidin arrangement for improved measurements of DNA hybridization kinetics. The latter is demonstrated by identifying a surface-coverage dependent viscoelastic behavior of immobilized b-DNA, which is shown to influence the hybridization efficiency.

Surface and Friction Behavior of a Silicone Surfactant Adsorbed on Model Textiles Substrates

“…34,35 The shifts in dissipation were <5% of the respective frequency shift. Additionally, little differences among the normalized overtone frequencies and dissipations were registered.…”

Specific and reversible DNA-directed self-assembly of oil-in-water emulsion droplets

“…This leads to a 1:1 ratio of DSPE-PEG-btn to btn-ssDNA oligonucleotides and consequently to a DNA surface coverage of about 15,200 strands/μm 2 . However, for btn-DNA coupled via streptavidin to a biotin-doped supported phospholipid bilayer, Larsson et al (20) showed that the intuitive assumption of a 2:1 molar ratio of btn-ssDNA oligonucleotides may be too simplistic and that the combination of electrostatic repulsion and steric hindrance may prevent efficient coupling of two btn-DNA oligonucleotides per streptavidin. In this case, only one btn-DNA oligonucleotide would be coupled per streptavidin; consequently, the DNA surface coverage would be halved.…”

Formation of supported bacterial lipid membrane mimics

“…After resolving the refractive index and optical thickness of the adsorbed layer, de Feijter's formula 45 was used to calculate the mass of adsorbed lipids using ͑dn / dc͒ lipid = 0.18. The value for dn / dc is not well defined in the literature and the value 0.18 cm 3 / g was chosen as an approximate median value found for planar lipid bilayers, 3,[46][47][48] where a higher value than for liposomes is expected due to alignment of the lipids in the evanescent field; the effective value could vary somewhat with lipid species.…”