Translational lysates form the foundation for the synthesis of proteins in cell-free systems. In order to ensure a correct translocation to the microsome membrane, endogenous microsomes in eukaryotic lysates are essential for the mapping of posttranslationally modified proteins, such as glycoproteins and membrane-bound proteins. The Eukaryotic Lysate Working Group has many years of experience in the cultivation of eukaryotic cell lines and their conversion to translational lysates for protein synthesis. Testing new cell lines for their capacity for in-vitro expression plays a large role. The development and continuous optimization of eukaryotic cell-free translation systems is an important research focus of the Working Group. The effect of fermentation and cell disruption as well as the transcription and translation components for the productivity of lysates are of key interest in this regard.