Hoffmann-la Roche Inc.

Scil Proteins Gmbh

F. Hoffmann-la Roche Ag

Rudolph, Rainer

Schwarz, Elisabeth

Elisabeth Schwarz

Rainer Rudolph

Adelbert Grossmann

Dorothee Ambrosius

Anke Rattenholl

Joerg Schaeffner

Gerhard Herr

Frank Hillger

Jörg Schaeffner

The invention relates to a method for producing biologically active &#x3b2;-NGF from the proform proNGF. After expressing the proform of the &#x3b2;-NGF in a prokaryotic host cell, the recombinant protein is isolated in the form of insoluble inactive aggregates (inclusion bodies). After the solubilization thereof in a strong denaturing agent and the subsequent conversion thereof into the natural conformation, which is determined by the disulfide bridges present in the natural &#x3b2;-NGF, biologically active &#x3b2;-NGF is obtained by subsequently splitting-off the prosequence.

Method for obtaining active pro-NGF and beta-NGF

Pharmaceutical preparations comprising human proNGF

A process for the production of a naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges by a) culturing prokaryotic cells in which the said prokaryotic cells contain an expression vector which codes for the said polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) secreting the polypeptide into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium, which is characterized in that a nucleic acid coding for a molecular chaperone is additionally expressed in the said prokaryotic cell and the chaperone is secreted into the periplasm, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.

Process for the production of naturally folded and secreted proteins by co-secretion of molecular chaperones

A process produces a water-soluble, naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges. This process involves culturing prokaryotic cells, a) in which the prokaryotic cells contain an expression vector which encodes the polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) under conditions under which the polypeptide is secreted into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium. In this process, the culturing is carried out in the presence of arginine or a compound of the formula I R,&mdash;CO&mdash;NR,(I) in which R and R,represent hydrogen or a saturated or unsaturated branched or unbranched C,-C,alkyl chain and R,represents hydrogen, NHR,or a saturated or unsaturated branched or unbranched C,-C,alkyl chain, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.

Process for the production of naturally folded and secreted proteins

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Halle, Germany

Elisabeth Schwarz