Nanodisc based Cell-free Membrane Protein Expression Service

As one of the best biological membrane-mimicking systems, nanodiscs are widely used in the solubilization and stabilization of membrane proteins. Membrane proteins incorporated in nanodiscs can be applied in immunization, drug and antibody screening, protein-protein interaction study using SPR and BLI, structural study using NMR and cryo-EM, etc. Therefore, production of membrane proteins in nanodiscs is in high demand nowadays. Experienced in both membrane protein expression and nanodisc preparation, Creative Biolabs now provides the high-quality custom service in cell-free membrane protein expression into nanodiscs.

Magic™ Cell-Free Membrane Protein Expressions into Nanodiscs

Compared with cell-based protein expression, one does not need to worry about the toxicity of expressed proteins to host cells, or insufficient membrane insertion, aggregation and degradation of target membrane proteins when membrane proteins are synthesized via cell-free protein expression. Besides, benefits of using cell-free protein expression systems also include the possibilities of chemical modification and unnatural amino acid incorporation in synthesized proteins.

Creative Biolabs provides the following innovative cell-free expression systems for this service.

Prokaryotic System	Cell-Free Protein Expression in E. coli Lysates
Eukaryotic Systems	Cell-Free Protein Expression in Wheat Germ Lysates Cell-Free Protein Expression in CHO Cell Lysates Cell-Free Protein Expression in Rabbit Reticulocyte Lysates Cell-Free Protein Expression in Insect Cell Lysates Cell-Free Protein Expression in HEK Cell Lysates
PURE System	Cell-Free Protein Expression in the PURE System

In this service, newly synthesized membrane proteins will be directly incorporated into pre-assembled nanodiscs using bilayer-forming lipids and membrane scaffold proteins (MSPs). NO DETERGENT will be used in any step. Besides, lipid-bilayer additives (e.g. sterols) may also be incorporated in nanodiscs to facilitate this process. The types of lipids and lipid-bilayer additives may greatly affect the conformation, oligomeric state, function, and activity of incorporated membrane proteins. We will also design and select the most matching MSPs according to different protein targets.

Magic™ Cell-Free Membrane Protein Expressions into Nanodiscs

In order to keep membrane proteins in their native-like format, screening of proper lipids, lipid-bilayer additives and MSPs is very critical.

First, based on our lipid screening platform, more than 30 bilayer-forming lipids with different tail lengths, saturation levels, and head group charges, various lipid-bilayer additives are available for screening.
Second, we also provide MSPs with multiple lengths to suit proteins with different sizes.
Third, we will also test the effect of different lipid-bilayer additives on membrane protein folding, conformation, oligomerization, function and activity according to custom requests.

During the comprehensive screening procedures, we will also test the compatibility of nanodiscs with every component in cell-free expression solutions so that the inclusion of nanodiscs will not impact protein expression efficiency.

In addition to the nanodisc format, we also provide alternative services of cell-free membrane protein synthesis and reconstitution into micelles and liposomes. Our skillful scientists in Creative Biolabs sincerely look forward to working with global researchers, providing professional assistance on their membrane protein programs. Please feel free to inquire us for more information.

Reference

Claxton, D.P.; et al. "Navigating Membrane Protein Structure, Dynamics, and Energy Landscapes Using Spin Labeling and EPR Spectroscopy." Methods in Enzymology. 2015, 564: 349-387.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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