Formation and cross-linking of fibrinogen layers monitored with in situ spectroscopic ellipsometry

Abstract

Thick matrices of fibrinogen with incorporation of a matrix metalloproteinase inhibitor were covalently bonded on functionalized silicon surfaces using an ethyl-3-dimethyl-aminopropyl-carbodiimide and N-hydroxy-succinimide affinity ligand coupling chemistry. The growth of the structure was followed in situ using dynamic ellipsometry and characterized at steady-state with spectroscopic ellipsometry. The growth was compared with earlier work on ex situ growth of fibrinogen layers studied by single wavelength ellipsometry. It is found that in situ growth and ex situ growth yield different structural properties of the formed protein matrix. Fibrinogen matrices with thicknesses up to 58 nm and surface mass densities of 1.6 μg/cm 2 have been produced.

Adsorption of Glucose Oxidase to 3‐D Scaffolds of Carbon Nanotubes: Analytical Applications

“…The resulting optical model is schematically shown in Figure 1B. In agreement with previous reports describing the adsorption of transparent polymers [55-57], the optical properties of the enzyme were described using a Cauchy model (A=1.45, B=0.1, C=0). This volumetric interpretation of the adsorption process is also coherent with previous reports [58, 59].…”

Yeast Cytochrome c Monolayer on Flat and Nanostructured Gold Films Studied by UV–Vis Spectroscopic Ellipsometry

“…Spectroscopic Ellipsometry Ellipsometry and UV-vis SE are often used to investigate adsorption dynamics at surfaces and thickness of thin and ultrathin biological films [22][23][24][25][26] and other complex molecular systems [7,8,[27][28][29][30][31].…”

Fibrinogen adsorption on blocked surface of albumin

“…Even though protein monolayer adsorption is a common hypothesis within the community of protein adsorption and biomaterials, there are also studies arguing for the possibility of proteins adsorbing in a multilayer fashion [9,15,[33][34][35]. Results from experiments with radioactive labelled proteins suggest that the hydrophobic PET surface can induce large conformational changes in adsorbed proteins and that those changes can induce enhanced adsorption of later arriving proteins onto the surface [24].…”

“…Results from experiments with radioactive labelled proteins suggest that the hydrophobic PET surface can induce large conformational changes in adsorbed proteins and that those changes can induce enhanced adsorption of later arriving proteins onto the surface [24]. Vogler et al [15,33,35,36] used what they call 'volumetric interpretation' to suggest a model for multilayer adsorption, and our results strengthens their idea of the existence of an interface region between surface and solution where the protein concentration can be orders of magnitude higher than in solution and where more or less ordered matrix structure of interacting proteins can be formed. In this study, the influence of pre-adsorption of albumin on fibrinogen adsorption onto PET and glass surfaces is investigated using a radioactive multi-label technique.…”