Acrylate-based hydrogels with multifunctional properties have proven to be suitable candidates for the development of sensor systems. They gained popularity especially in combination with bioelectronics, as there is a need to understand and control the interactions of bionic devices with the human body and other environments. In this study, we present results on the biointeraction capability of plasma-polymerized (pp) hydrogels made of hydroxyethyl methacrylate (HEMA) and 2-(diethylamino)ethyl methacrylate (DEAEMA) mixtures on gold screen-printed electrodes (SPE). The hydrogels were generated by an atmospheric pressure plasma jet, and their chemical composition was characterized via FT-IR. The FT-IR analysis revealed several functional groups suitable for biomolecule immobilization, whereas the amount of -C-N, –OH, and -C-O-C groups differs depending on the mixture ratios. The pp HEMA:DEAEMA (HD) hydrogel coatings provide alternative interfacing materials for electrochemical biosensing. The enzymes glucose oxidase (GOx) and acetylcholinesterase (AChE) were coupled to the hydrogel-based surfaces, and the effects of the mixture ratios on the biomolecule immobilization were investigated. It is possible to address different functional groups of the mixtures with different immobilization strategies; thus, the sensor response can be optimized. Finally, glucose as GOx substrate and eserine as AChE inhibitor were detected by amperometry to demonstrate the practical biosensing applicability of the coatings.