Separation of wheat flour into its constituents starch and gluten was studied using a cone-cone shearing device, with emphasis on the effect of rotation rate, processing time, temperature and water content. This study confirms the two step mechanism previously proposed for the gluten migration: aggregation of gluten protein into gluten domains that subsequently migrate to the apex of the cone. The results show that optimal process conditions for gluten migration are different from the process conditions for gluten aggregation. While gluten agglomeration (step 1) benefits from high temperature, low rotation rate and high water content, gluten migration (step 2) is positively influenced by a high dough viscosity and higher rotation rate. (C) 2009 Elsevier Ltd. All rights reserved.