Specific Protein Binding to Functionalized Interfaces

Schmitt, Franz Josef; Blankenburg, R.; Häußling, Lukas; Ringsdorf, Helmut; Weisenhorn, A. L.; Hansma, Paul K.; Leckband, Deborah; Israelachvili, Jacob N.; Knoll, Wolfgang

doi:10.1007/978-1-4899-1630-3_12

Cited by 3 publications

(2 citation statements)

References 25 publications

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“…The shift in the surface plasmon peak, obtained before and after SAV incubation, was used to calculate the thickness d of the SAV layer (Winspall software, from RES-TEC, Germany). The model was simplified by taking the biotinylated PMMA as a 52-nm-thick single layer with a refractive index of 1.50. , From thickness results and considering a theoretical thickness value of 5 nm and a refractive index of 1.45 for a homogeneous monolayer of SAV, , the surface coverage can be calculated. Also, SAV surface density A can be calculated using eq A = d false( n − n 0 false) ∂ n / ∂ c where A is the adsorbed amount of protein per area and n 0 is the refractive index of the solution and with ∂ n /∂ c = 0.212 mL/g (derived independently by Knoll et al from refractometric measurements) …”

Section: Methodsmentioning

confidence: 99%

Universal Chemical Gradient Platforms Using Poly(methyl methacrylate) Based on the Biotin−Streptavidin Interaction for Biological Applications

et al. 2010

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This article describes a simple method for the construction of a universal surface chemical gradient platform based on the biotin-streptavidin model. In this approach, surface chemical gradients were prepared in poly(methyl methacrylate) (PMMA), a biocompatible polymer, by a controlled hydrolysis procedure. The physicochemical properties of the resulting modified surfaces were extensively characterized. Chemical analysis carried out via time-of-flight secondary ion mass spectrometry (ToF-SIMS) and X-ray photoelectron spectroscopy (XPS) showed the formation of a smooth, highly controllable carboxylic acid gradient of increasing concentration along the sample surface. Atomic force microscopy (AFM) and contact angle (CA) results indicate that, in contrast with most of the chemical gradient methods published in the literature, the chemical modification of the polymer surface barely affects its physical properties. The introduction of carboxylic acid functionality along the surface was then used for biomolecule anchoring. For this purpose, the surface was activated and derivatized first with biotin and finally with streptavidin (SAV) in a directed orientation fashion. The SAV gradient was qualitatively assessed by fluorescence microscopy analysis and quantified by surface plasmon resonance (SPR) in order to establish a quantitative relationship between SAV surface densities and the surface location. The usefulness of the fabrication method described for biological applications was tested by immobilizing biotinylated bradykinin onto the SAV gradient. This proof-of-concept application shows the effectiveness of the concentration range of the gradient because the effects of bradykinin on cell morphology were observed to increase gradually with increasing drug concentrations. The intrinsic characteristics of the fabricated gradient platform (absence of physicochemical modifications other than those due to the biomolecules included) allow us to attribute cell behavior unequivocally to the biomolecule surface density changes.

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Section: Methodsmentioning

confidence: 99%

Universal Chemical Gradient Platforms Using Poly(methyl methacrylate) Based on the Biotin−Streptavidin Interaction for Biological Applications

et al. 2010

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“…It has been shown that these interfaces can have a high binding affinity for streptavidin ifone carefully engineers the molecular structure, the site density, relative orientational constraints, etc. (12).…”

Section: Introductionmentioning

confidence: 99%

Streptavidin binding to biotinylated lipid layers on solid supports. A neutron reflection and surface plasmon optical study

Schmidt

Spinke

Bayerl

et al. 1992

Biophysical Journal

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Neutron reflection and surface plasmon optical experiments have been performed to evaluate structural data of the interfacial binding reaction between the protein streptavidin and a solid-supported lipid monolayer partly functionalized by biotin moieties. Since both experimental techniques operate in a total internal reflection geometry at a substrate/solution interface, identical sample architectures allow for a direct comparison between the results obtained with these two recently developed methods. It is found that a monomolecular layer of dipalmitoyllecithin doped with 5 mol% of a biotinylated-phosphatidylethanolamine shows a thickness of d1 approximately (3.4 +/- 0.5) nm. Binding of streptavidin to the biotin groups results in an overall layer thickness of d = (5.9 + 0.5) nm that demonstrates the formation of a well-ordered protein monolayer with the (biotin+spacer) units of the functionalized lipids being fully embedded into the binding pocket of the proteins. It is demonstrated by model calculations that a more detailed picture of the internal structure of this supramolecular assembly can only be obtained if one uses deuterated lipid molecules, thus generating a high contrast between individual layers.

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A new kind of immobilized enzyme multilayer based on cationic and anionic interaction

Kong¹,

Zhang²,

Gao³

et al. 1994

Macromol. Rapid Commun.

109

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Specific Protein Binding to Functionalized Interfaces

Cited by 3 publications

References 25 publications

Universal Chemical Gradient Platforms Using Poly(methyl methacrylate) Based on the Biotin−Streptavidin Interaction for Biological Applications

Universal Chemical Gradient Platforms Using Poly(methyl methacrylate) Based on the Biotin−Streptavidin Interaction for Biological Applications

Streptavidin binding to biotinylated lipid layers on solid supports. A neutron reflection and surface plasmon optical study

A new kind of immobilized enzyme multilayer based on cationic and anionic interaction

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