Ultrasensitive Fluorescence Microscopy Studies of Protein Interactions with Functionalized Surfaces
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Colin D. Heyes
Controlling protein adsorption to surfaces is an important issue in biotechnology, biomedicine and basic sciences. Proteins readily adsorb non-specifically to a variety of surfaces in a process known as biofouling, which is a major problem in the biomedical, environmental and industrial fields that can be solved by pre-treating the surfaces with adsorption-resistant materials. Additionally, surfaces can be structured and functionalized so as to immobilize proteins in specific patterns, orientations and/or densities, which is important in such areas as biosensors, biocatalysis, biotechnology, protein microarrays and single molecule biophysics. Here we review the various surface modifications and bioconjugation techniques that have been used in these areas. We also introduce novel fluorescence microscopy techniques, which have been employed to analyze the resistance of the surfaces to non-specific adsorption with detection limits down to the single molecule. A huge advantageous property of fluorescence microscopy is its sensitivity to the structure of individual, immobilized proteins by using Förster resonance energy transfer (FRET). Thus one can examine if the surface coating maintains the folded, functional form of specifically immobilized proteins. We then describe several important advances in studying slow timescale biophysical processes such as protein folding of single, specifically immobilized proteins. Future directions into which this field is expected to progress are also discussed.
© Oldenbourg Wissenschaftsverlag
Articles in the same Issue
- Bunsen Colloquium: Controlling Protein Adsorption at Materials Surfaces
- Adsorption of Collagen Fragments on Titanium Oxide Surfaces: A Molecular Dynamics Study
- Adsorption of the Flexible Salivary Proteins Statherin and PRP-1 to Negatively Charged Surfaces – A Monte Carlo Simulation and Ellipsometric Study
- Surface-Tethered Polymers to Influence Protein Adsorption and Microbial Adhesion
- Lactoperoxidase and Histatin 5 – their Adsorption Behaviour on Silica and Hydrophobized Silica Surfaces, and Implications on their Role in the Initial Salivary Film Formation
- Ultrasensitive Fluorescence Microscopy Studies of Protein Interactions with Functionalized Surfaces
- Adsorption of Amyloid β (1-40) Peptide at Liquid Interfaces
- Controlled Release of Proteins Bound to Spherical Polyelectrolyte Brushes
- In-Situ-ATR-FTIR Detection of Protein Sorption at Polyelectrolyte Multilayers: Variation of the Thickness
- Stability of Proteins Confined in MCM-48 Mesoporous Molecular Sieves – The Effects of pH, Temperature and Co-solvents
- Hydrogen Bonding of Water Confined in Controlled-Pore Glass 10-75 Studied by 1H-Solid State NMR
Articles in the same Issue
- Bunsen Colloquium: Controlling Protein Adsorption at Materials Surfaces
- Adsorption of Collagen Fragments on Titanium Oxide Surfaces: A Molecular Dynamics Study
- Adsorption of the Flexible Salivary Proteins Statherin and PRP-1 to Negatively Charged Surfaces – A Monte Carlo Simulation and Ellipsometric Study
- Surface-Tethered Polymers to Influence Protein Adsorption and Microbial Adhesion
- Lactoperoxidase and Histatin 5 – their Adsorption Behaviour on Silica and Hydrophobized Silica Surfaces, and Implications on their Role in the Initial Salivary Film Formation
- Ultrasensitive Fluorescence Microscopy Studies of Protein Interactions with Functionalized Surfaces
- Adsorption of Amyloid β (1-40) Peptide at Liquid Interfaces
- Controlled Release of Proteins Bound to Spherical Polyelectrolyte Brushes
- In-Situ-ATR-FTIR Detection of Protein Sorption at Polyelectrolyte Multilayers: Variation of the Thickness
- Stability of Proteins Confined in MCM-48 Mesoporous Molecular Sieves – The Effects of pH, Temperature and Co-solvents
- Hydrogen Bonding of Water Confined in Controlled-Pore Glass 10-75 Studied by 1H-Solid State NMR
