Fibrin glue is a two-component adhesive used to stop bleeding, seal wound edges and for scaffolds in tissue engineering. Autologous products result in reduced risk of contamination and immunological responses compared to commercially available fibrin glue. However, reproducibility due to patient dependent sealant properties of autologous fibrin glue preparation is low. In this study a fully automated production process for both the fibrinogen and thrombin component from small blood volumes was developed. The resulting fibrinogen concentration, thrombin activity and sealant properties of the fibrin glue were determined. The fully automated and closed production system proved to be a promising tool for fast and easy production of autologous fibrin glue with an effective adhesive quality.