Large-scale recombinant proteins in Gram-negative bacteria (i.e., E. coli) are of significant interest in recombinant protein/enzyme production. These proteins exhibit wide-ranging applications, including food and beverages, conversion of carbohydrates into fuel ethanol or biodiesel, components for clothing and cosmetics, biopolymers, cleaning materials, and waste management.

The Need
Although there are well-established protocols for the purification of recombinant proteins, complex and expensive purification methods limit the large-scale production of these proteins. Chromatography is the leading purification modality, especially in the biotechnology and pharmaceutical industry, but it is impractical in many other areas. New purification techniques are needed to facilitate the broad use of recombinant proteins.

The Technology
This technology describes a novel high-throughput recombinant protein purification method that can be used for large-scale production. The technology allows a protein of interest is fused to a sequence to engender formation of fibrils, allowing separation from other cellular proteins. The protein of interest is then released in soluble form without a chromatographic step. As proof of concept for this fibril-mediated purification we isolated a bacterial phosphatase without the use of any chromatographic step. The fibril-forming sequence can be removed through various approaches, thus affording the recombinant protein without any additional sequence.

Commercial Applications
This invention can be used to purify recombinant proteins for several industries, including but not limited to pharmaceuticals, agriculture, biopolymers, biofuels, and others.

Benefits/Advantages
Compared to chromatography, this invention provides a straightforward, easy-to-use, and inexpensive method for the large-scale purification and production of proteins.

Patents
Provisional patent application filed.