Patent ID: 7351549

Claim:
A method for the manufacture of recombinant trypsinogen or trypsin comprising incubating transformed host cells expressing a trypsinogen gene in a suitable cell culture nutrient medium at conditions allowing for cell multiplication and simultaneous or subsequent expression of the trypsinogen gene, separating extracellularly accumulated trypsinogen from the cell suspension, characterized in that the method comprises incubating the transformed host cells at an acidic pH at or above pH 2, the transformed host cells being yeast cells transformed with an expression vector that contains a naturally occurring trypsinogen gene, and after incubation, during which no trypsinogen activation or trypsin activity is detected in the medium by SDS polyacrylamide gel electrophoresis, separating the cells from the medium, obtaining a solid-free supernatant containing correctly folded trypsinogen and subjecting said solid-free supernatant containing correctly folded trypsinogen to purification by chromatography without a preceding refolding or buffer exchanging step, and optionally further processing the separated trypsinogen.