Method of Producing Soluble Recombinant Proteins from Bacterial Expression Systems

Background:

Recombinant proteins are encoded by recombinant DNA that has been cloned in an expression vector that supports expression of the gene and translation of messenger RNA. They are generated in various ways to increase production of proteins, modify gene sequences, and manufacture a wide variety of commercial products. Recombinant proteins that are synthesized in bacterial expression systems are frequently found in inclusion bodies, and are therefore not soluble. As a result, they form precipitates and are functionally inactive in aqueous solutions, limiting their usability for laboratory research. 

Technology Overview:

This technology is a method for producing soluble proteins that are normally insoluble in bacterial expression systems. This enables the production of functionally active recombinant proteins in a way that saves significant time and effort. To do this, a recombinant protein is produced in a bacterial expression system that includes an affinity tab on the protein’s C and/or N terminus. The bacteria are then lysed and a precipitate of the protein forms. The protein is washed and 8M urea is added to solubilize the precipitate. Next, the recombinant protein is purified and eluted with an imidazole/urea buffer. The preparation is then dialyzed in a urea borate buffer to remove imidazole, and treated with 3.0 mM NHD biotin followed by dialysis with an NaCl/borate buffer to remove urea and free biotin. This results in soluble biotinylated recombinant protein.

Advantages:

•    Produces soluble recombinant proteins from bacterial expression systems.
•    Produces more proteins than eukaryotic expression methods.
•    Saves time and effort.

Applications:

The technology’s primary application is to produce recombinant proteins for biological research.

Intellectual Property Summary:

Patent application filed: PCT/US2021/062280

Stage of Development:

TRL 3 - Experimental proof of concept

Licensing Status:

This technology is available for licensing.

Licensing Potential:

This technology would be of interest to anyone involved in research involving recombinant proteins. This includes:
•    University laboratories.
•    Medical facilities.
•    Pharmaceutical laboratories.