Besides providing cGMP, preclinical and protein engineering services, a key area that Protein Technologies Ltd dedicates itself to is the design and construction of new biological parts, devices, and systems that provide useful purposes now and in the future. We have used our expertise in protein engineering and to produce and offer (i) the next generation of proteins and (ii) Ice-crystal biosensors.
The next generation of proteins (unnatural proteins)
By manipulating the protein production machinery in bacteria we can incorporate unnatural elements such as fluorine not found in naturally occurring proteins to manufacture biosynthetic molecules at scale.
Proteins manufactured in this way can be used in applications in bio-therapeutics and diagnostics and the production of vaccines as they can provide increased membrane permeability, longevity and stability and greater potency.
In most industrial fermentations, yield is the most critical determinant of success, being the primary factor affecting cost. Here we are much more concerned with establishing levels of unnatural amino acid incorporation.
Levels of 95% incorporation can be achieved (results verified by mass spec and NMR) demonstrating that the process is indeed fully scalable.
An unnatural analogue of Green Fluorescent Protein produced at Protein Technologies Ltd referred to as Kryptonite (Figure 1) with an extra peak at 475 nm (Figure 2). Note that this additional peak is much larger in the fluorinated protein and subsequent fluorescence, at 475 nm excitation, is much larger than the wild-type protein. Maybe figure showing 19F NMR.
By genetically modifying fluorescent protein and incubating them in a milieu of specific agents we have produced biosensors for the detection and reporting of ice-crystal formation.
These fluorescent proteins have been engineered to be hypersensitive to the formation of large ice-crystals. Normally the protein fluoresces a bright red/green colour, however, when it is exposed to the slow freezing conditions in which large crystals are formed, it is irreversibly denatured and turns an opaque white.
These proteins are the ideal reagents to indicate the deleterious effects of ice-crystal formation on the structural integrity of the proteins and proteinaceous compounds of which biological samples and products are themselves composed.
Our ice-crystal biosensors exhibit a number of further advantages over conventional freeze detection solutions. As a liquid suspension, they lend themselves to formulations in blister packs or small stick-on labels; traditional freeze-sensitive labels and eTTI’s are far too bulky to be attached to individual products.
A second major advantage is cost. The colloid-filled membranes on which freeze-sensitive labels are based call for complex fabrication and each eTTI requires a separate electronic circuit board. Because our ice-crystal biosensors are produced by fermentation, it can be manufactured as a bulk liquid at low cost.