Microbiological investigations are the state-of-the-art method to evaluate aseptic sterilization machines. Recently, different sensors have been introduced to overcome these time-consuming and lab-intensive procedures, for example, calorimetric sensors or spore-based biosensors. The former are developed from microbiological spores, which have to be properly immobilized on the sensor's surface to provide a reliable and consistent sensor signal in order to evaluate sterilization processes with gaseous hydrogen peroxide. Still, no stable, oxidation-resistant and harmless immobilization technique exists for the immobilization of bacterial spores. The aim of this work is, therefore, to elaborate an oxidation-resistant functionalization method to immobilize B. atrophaeus spores for spore-based biosensors. (C) 2019 Elsevier Ltd. All rights reserved.