Wheat gluten-chitosan composite film (WGCCF) can prevent moisture migration and enhance the antimicrobial properties of gluten in intermediate-moisture foods like sandwiches. To mimic the structure of actual sandwich-type products we developed multi-layer food models, where moisture content and water activity differ. Water activity gradients direct moisture migration and therefore determine product characteristics and product stability. A 10% wheat gluten film-forming solution was mixed with chitosan film-forming solution (0-3%, w/w) and evaporated to generate WGCCF. Addition of 3% chitosan enhanced the mechanical properties of the film composite, lowered its water vapor permeability, and improved its ability to protect against both, Streptococcus faecalis and Escherichia coli, in a 24 hr sandwich test (reduction of 1.3 and 2.7 log cycles, respectively, compared to controls). Best barrier and antimicrobial performance was found for 3% chitosan WGCCF at pH 5.1. Film of this type may find application as barrier film for intermediate-moisture foods.