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The invention relates to methods for obtaining and purifying therapeutic polypeptides, particularly polypeptides produced in recombinant microbial hosts. The invention further relates to compositions and methods for therapeutic uses of such polypeptides.
Polypeptides have become important therapeutic agents in a wide variety of illnesses and disorders. Administration of polypeptides can take a number of different forms, including oral and parenteral delivery to patients. The delivery of polypeptides, however, often requires that they first be synthesized by some suitable method. For polypeptides which are produced in microbial hosts, the traditional way of producing therapeutically effective amounts of such polypeptides has been to purify them from the culture medium. This has required initial growth of the bacteria in a suitable nutrient medium, in the presence of compounds such as antibiotics to produce a suitable culture medium, followed by purification of the polypeptide.
This traditional method has several shortcomings. It is limited by the available purification techniques to polypeptides with molecular weight less than about 50,000. It requires specialized instrumentation, e.g., a high pressure centrifuge or chromatographic resins. The various purification techniques do not generally separate polypeptides with molecular weights greater than about 10,000 efficiently, and so many of the polypeptides in a culture supernatant, after separation from cells, are lost for further use. Finally, the purification process frequently disrupts the polypeptide so that it is no longer biologically active.
An alternate method for obtaining and purifying polypeptides is disclosed in U.S. Pat. No. 4,431,739 (WO 83/02755). The disclosure and claims are directed to a method for recovering a bacterial protein which differs from a native protein in its amino acid sequence. In the described method, a high level, heterologous expression host cell is transformed with an expression vector which contains within its DNA sequence a portion of a gene encoding the bacterial protein in frame with the
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