Phage Biotechnology Corp.

Technology

Technology | Manufacturing Process

Technology

Phage has developed a proprietary manufacturing process, covered by numerous U.S. and foreign patents, to produce protein pharmaceutical products. This process involves the synthesis, at high levels, of recombinant proteins in the bacterium, E. Coli. Unlike existing plasmid technologies, where the total intracellular recombinant protein product is generally produced as an insoluble, unfolded, and inactive product, the Phage process produces a protein product that is soluble, folded properly, and in a biologically active state. This fact greatly simplifies the purification of the protein product and results in a biological product that has not been subjected to harsh chemical treatments. The simplified purification process leads to a more efficient and rapid manufacturing process.

In addition, Phage has partnered with BiopolyMed, Inc., a Korean biotechnology company that has developed a proprietary method to pegylate protein pharmaceuticals. This technology will be applied to Phage products where a longer acting version of the drug (for example, peg-growth hormone, peg-interferon alpha 2b) will greatly expand the market for these products.

Phage Manufacturing Process

The ability of the Phage method to produce biologically active, soluble recombinant proteins within bacteria gives the company a significant production advantage over other microbial methods of recombinant drug manufacturing. Denaturation and solubilization of the recombinant protein from inclusion bodies is avoided in the Phage process, as well as the lengthy and relatively inefficient refolding of the protein, where as much as 90% of the recombinant protein can be misfolded and must be discarded. This fact greatly simplifies the purification of the protein product and results in a biological product that has not been subjected to harsh chemical treatments.
Briefly, the manufacturing process involves the cloning of a human gene sequence (for example, human interferon or human growth hormone) into a bacterial plasmid, which is then introduced into the bacteria. At a defined moment in time the bacteria are infected with bacteriophage, which results in the super-synthesis of the desired human product, followed by lysis of the bacteria liberating the human product in a soluble, completely active form. The desired protein product is then purified to purity using conventional chromatographic procedures. This process has been utilized for more than 8 years to manufacture numerous proteins of human, animal or viral origin. It is a simple and highly cost-efficient method to manufacture proteins of commercial value.
The two figures below conceptualize the initial state of the recombinant protein product in either the phage or plasmid method. In the Phage process the protein product appears in the interior of the cell already folded (ribbon-like structures in the figure). In the plasmid procedure the protein products appear as insoluble inclusion bodies depicted as solid balls in the figure below. These inclusion bodies must first be solubilized in strong denaturants and the product re-folded, steps that add cost and time to the purification process, and in some cases, can damage the protein product.

Phage Process		Plasmid Process

We have done in-house comparisons of the costs and time involved in producing recombinant proteins inside bacteria by the Phage versus Plasmid method. We estimate that, per gram of recombinant protein manufactured, the Phage process has a shorter run time for each production cycle of nearly 50% (due largely to the time required for the refolding step necessary in the Plasmid procedure). In addition, since the refolding step is highly inefficient, our cost of producing a gram of biologically active recombinant protein is substantially reduced when compared to the Plasmid method where inclusion bodies are formed.

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