Creative BioMart is a well-known expert specializing in the application of advanced cell-free display technology to accelerate the release of protein engineering. With years of experience, we are able to provide in vitro compartmentalization display system service based on cell-free technology to precisely meet customer requirements.

The in vitro compartmentalization is a powerful tool that can be used as protein engineering because it combines the flexibility of plate-based detection with larger libraries and quantitative high-throughput screening techniques (such as flow cytometry). The system is based on water-in-oil (W/O) emulsion droplets as cell-like compartments. In each drop, a single gene is transcribed and translated to provide multiple copies of the protein it encodes. This ensures that the gene, the protein it encodes, and the product of the protein's activity are kept in the same compartment, thereby providing a link between the gene and its molecular phenotype. By applying appropriate selection pressure, genes encoding proteins with the desired activity (binding or enzymatic) can be selected from a large gene pool.

Services

Creative BioMart provides a complete in vitro compartmentalization display system based on emulsion technology. In this system, you can select bacterial cells, wheat germ, rabbit reticulocyte (RRL) extracts for transcription and translation, and stir the aqueous solution of gene and the in vitro transcription-translation system (covalent DNA display and noncovalent DNA display) to oil-surface activity to produce a water-in-oil (w/o) emulsion with 10¹⁰ water droplets per milliliter of emulsion. In addition, we use a fluorescence-activated cell sorter to convert w/o emulsions into water-in-oil-in-water (w/o/w) emulsions for high-throughput screening, allowing in the case of water droplets, an external water phase is produced.

The in vitro compartmentalization display system we provide is used to generate "artificial cells" and directed evolution of proteins.

Fig 1. Schematic representation of directed evolution by in vitro compartmentalization. (Miller OJ, et al., 2006)

Features of In Vitro Compartmentalization Display System

• With a bead display selection strategy based on in vitro compartmentalization, which is amenable to screening libraries of up to 10¹² protein variants by allowing beads to be overloaded with up to 10⁴ unique mutants.
• In vitro compartmentalization system can produce droplets with a diameter of 1-50 um. The formation and destruction of the emulsion takes 5-10 minutes, and only basic laboratory equipment is required. The emulsion can remain highly stable for several days or even months even at a temperature close to 100°C.
• In vitro compartmentalization can control the reaction within the droplet. The hydrophobic and hydrophilic components can be delivered to each droplet in a stepwise manner without compromising the chemical integrity of the droplet.
• In vitro compartmentalization allows the selection of catalytic molecules, and can choose to use anti-product antibodies and flow cytometry to detect product formation and quantity.

Creative BioMart has been continuously developing cutting-edge cell-free display technology, and is committed to providing a flexible and efficient customized in vitro compartmentalization display system service. Based on the protein engineering platform, we have successfully constructed an impeccable cell-free surface display library of various types. We are proud to be able to screen and select proteins with the required characteristics from a large number of variants to best accelerate the research of life sciences. If you are interested in our services, please do not hesitate to contact us for more information.

References

1. Miller OJ, Bernath K, et al.. (2006) Directed evolution by in vitro compartmentalization. Nat Methods. 3(7): 561-570.
2. Bernath K, Magdassi S, Tawfik DS. (2004) In vitro compartmentalization (IVC): A high-throughput screening technology using emulsions and FACS. Discov Med. 4(20): 49-53.

• Covalent DNA Displays Service
• mRNA Displays Service
• Noncovalent DNA Displays Service
• Ribosome Displays Service