Roche Diagnostics Operations, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-La Roche Ag

Hellmut Eckstein

Johann-Peter Thalhofer

Frank Geipel

Stephan Glaser

Claus S Schneider

Werner Hoelke

Rainer Muller

Stephanie Ronning

Bernhard Rexer

Michael Ratka

Claudia Giessel

Rainer Mueller

Bernhard Suppmann

Bettina Bommarius

Thomas Kirschbaum

Antje Liehre

Michaela Fischer

Claus Schneider

Elvira Wolf

The present invention provides a method for purifyingcollagenase type I and type II proteins from a complex mixture by subsequently performing a precipitation with ammonium sulfate, hydrophobic interaction chromatography, cation exchange chromatography, and anion chromatography. Conditions are provided which lead to a stabilized, partially purified preparation even after the precipitation step. The method of the invention leads to a quick and efficient removal of other proteolytic activities. The preparations according to the invention provide exceptionally pure and intact collagenase type I and type II proteins which are enzymatically active. The invention also provides blends of the two isolated proteins. The invention further provides the use of the purified collagenase proteins or blends thereof for treating a tissue sample in vitro.

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The present invention concerns a method for producing recombinant trypsin from porcine pancreas inwhich is soluble and secreted into the culture medium, whereby expression at pH 3.0-4.0 substantially prevents activation of trypsinogen to &#x3b2;-trypsin and autolysis of &#x3b2;-trypsin by &#x3b1;-trypsin into &#x3b5;-trypsin and from there into inactive peptides.

Method for producing recombinant trypsin

This invention involves a process for selecting and producing eukaryotic alkaline phosphatase in yeast. Yeast cells are subjected to multiple transformations using a vector comprising a first resistance marker gene and the alkaline phosphatase gene. Those strains that grow in media containing the first resistance marker are further transformed using a vector comprising a second selection marker gene and the alkaline phosphatase gene. Transformants that grow in media containing the second selection marker are selected for expressing the eukaryotic alkaline phosphatase.

Expression of alkaline phosphatase in yeast

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Weilheim, Germany

Hellmut Eckstein