METHOD FOR THE PREPARATION OF RECOMBINANT MAMMALIAN HEPARIN-BINDING PROTEIN (HBP)

摘要

The present invention relates to methods of making heparin-binding protein (HBP) in a recombinant bacterial expression system. In particular, this invention relates to a method for the preparation of an insoluble fusion protein comprising heparin-binding protein (HBP), a proteolytic cleavage- site, and a second polypeptide in recombinant bacterial cells, said fusion protein being accumulated in inclusion bodies in the cytoplasm of bacterium after expression. The invention further relates to methods of separation of the expressed HBP from inclusion bodies comprising purification of HBP from the second polypeptide and optionally refolding of said HBP. The invention further features the DNA constructs comprising different HBP-fusion proteins. Invention also relates to use of the HBP-fusion protein produced in bacteria for the production of pure HBP and use of the HBP purified from the fusion protein for the preparation of a medicament.