Process for producing recombinant trypsin

A process for refolding recombinant trypsinogen produced from a prokaryote host cell comprising:

• (a) providing the recombinant trypsinogen at a concentration of from 5 to 15 g/L in a solubilization solution comprising a chaotropic agent, a first buffer agent, and a first low molecular weight reducing agent, wherein the solubilization solution does not include a low molecular weight oxidizing agent that would provide a redox pair with the first low molecular weight reducing agent;
• and
• (b) infusing the solubilization solution comprising the recombinant trypsinogen of step (a) over time into a diluent comprising a second buffer agent, a second low molecular weight reducing agent, the chaotropic agent, and arginine to provide a refold solution that comprises the recombinant trypsinogen at a concentration greater than 1 g/L, the chaotropic agent at from 20% to 30% of its concentration in the solubilization solution of step (a), the first and second buffer agents, arginine, and the first and second low molecular weight reducing agents and incubating the refold solution for a time sufficient for the recombinant trypsinogen to refold into a conformation characteristic of native trypsinogen and form the disulfide bonds characteristic of native trypsinogen, wherein the refold solution does not include a low molecular weight oxidizing agent that would provide a redox pair with either the first or second low molecular weight reducing agents.