Detergent Micelle Platform

Proteins are inserted in lipid bilayers through hydrophobic interactions between lipid tails and hydrophobic protein structural domains. These integral membrane proteins are insoluble in aqueous solution and are aggregated. However, they can be soluble in descaling agents, which form micelles similar to the bilayer of a biological membrane. Proteins are encapsulated in these micelles through hydrophobic interactions. The hydrophobic regions of membrane proteins typically embedded in membrane lipid bilayers are surrounded by a layer of descaling agent molecules, while the hydrophilic regions are exposed to the aqueous medium.

Figure 1. Workflow of membrane protein purification mediated by detergents (Eberhard, C., 2012).

Detergent technology can be applied to a variety of expression systems, including bacterial, yeast, insect cell, mammalian cell, and cell-free expression systems. Each expression system has its own unique advantages and disadvantages and needs to be selected based on actual needs.

Membrane protein extraction process:

• Dissolve the membrane proteins using a suitable detergent.
• Initial purification using an ion exchange chromatography column to collect the target protein eluate.
• Concentrate for the final step of purification and further separate using a high-resolution chromatography column to obtain high homogeneity and high purity membrane proteins for subsequent analysis.

Types of Detergents	Advantages	Disadvantages
Nonionic detergent	Mild and less likely to denature proteins	Lower dissolution yields
Ionic detergents (cationic and anionic)	Effective in solubilizing membrane proteins	Causes protein denaturation and interference with ion exchange chromatography
Amphoteric ionic detergents	Combines the advantages of ionic and nonionic detergents	More likely to cause protein denaturation than nonionic detergents

Creative Biomart's experts design experiments based on the characteristics of the target protein, including screening of detergents, combinations of detergents, changes in extraction time, temperature, and method, screening of additives, and selection of purification methods (affinity chromatography, ion exchange chromatography, gel filtration/size exclusion, etc.). Gel-based or gel-free quality control for high-throughput analysis ensures that the final product meets customer standards.

Creative Biomart has accumulated extensive experience in membrane protein separation and purification. Our experienced scientists, proven technology platforms, and advanced facilities guarantee our high-quality services. Our competitive prices and rich expertise have earned us the trust of our collaborators. If you have any questions, please contact us.

Reference:

1. Eberhard, C. (2012). Development of a model system to study cell adhesion and cell mechanics (Doctoral dissertation).