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PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
Published on: July 27, 2017
Protein adsorption on supported phospholipid bilayers
Karin Glasmästar1, Charlotte Larsson, Fredrik Höök
1Department of Applied Physics, Chalmers University of Technology and Göteborg University, SE-412 96 Göteborg, Sweden. karing@fy.chalmers.se
Insights
Supported lipid bilayers made of egg-phosphatidylcholine (eggPC) lipids demonstrate significant protein resistance. Quartz crystal microbalance with dissipation (QCM-D) and surface plasmon resonance (SPR) confirmed minimal protein adsorption for various biomolecules.
Area of Science:
- Biomaterials Science
- Surface Chemistry
- Biophysics
Background:
- Supported lipid bilayers (SLBs) are crucial for biomimetic interfaces.
- Understanding protein adsorption onto SLBs is vital for biosensor and biomedical applications.
- Egg-phosphatidylcholine (eggPC) is a common lipid for forming stable SLBs.
Purpose of the Study:
- To investigate the protein resistance of supported eggPC lipid bilayers.
- To quantify protein adsorption using Quartz Crystal Microbalance with Dissipation (QCM-D) and Surface Plasmon Resonance (SPR).
- To compare protein adsorption of various biomolecules including fibrinogen, albumin, hemoglobin, cytochrome c, and immunoglobulin G (IgG).
Main Methods:
- Formation of supported eggPC lipid bilayers on SiO(2) surfaces via vesicle adhesion and fusion.
- Adsorption measurements using QCM-D for multiple proteins and 10% fetal bovine serum.
- Comparative adsorption studies using SPR for fibrinogen and hIgG.
Main Results:
- EggPC lipid bilayers exhibited high protein resistance, with irreversible adsorption below detection limits.
- Reversible protein adsorption was observed, ranging from 0.3-4% of saturation coverage on a gold thiol monolayer.
- SPR measurements indicated slightly higher adsorbed amounts compared to QCM-D.
Conclusions:
- Supported eggPC lipid bilayers demonstrate excellent protein-repellent properties.
- The findings suggest potential for using eggPC bilayers in applications requiring resistance to non-specific protein binding.
- Further investigation into the mechanisms underlying this protein resistance is warranted.
Abstract:
Quartz crystal microbalance with dissipation (QCM-D) measurements were used to investigate the adsorption of human fibrinogen, human serum albumin, bovine hemoglobin, horse heart cytochrome c, human immunoglobulin (hIgG), and 10% fetal bovine serum on supported bilayers of egg-phosphatidylcholine (eggPC) lipids. For comparison the adsorption of fibrinogen and hIgG to eggPC bilayers was also studied with surface plasmon resonance (SPR). The supported bilayers were formed in situ by vesicle adhesion and spontaneous fusion onto a SiO(2) surface. The supported lipid bilayer is highly protein resistant: The irreversible adsorption measured with the QCM-D technique was below the detection level, while reversible protein adsorption was detected for all the proteins in the range 0.3-4% of the saturation coverage on a hydrophobic thiol monolayer on gold. The adsorbed amounts were slightly higher for the SPR measurements. Possible mechanisms for the protein resistance of eggPC bilayers are briefly discussed.
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