https://pages.htgaa.org/2026a/violeta-vilcapoma-torres/homework/week-09-hw-cell-free-systems/index.html1. Advantages of CFPS The main advantage is that CFPS is an open system. Unlike in vivo methods, there is no cell membrane, allowing direct access to the reaction. Flexibility: You can adjust $Mg^{2+}$ levels, add chaperones, or use non-natural amino acids easily. Toxic Proteins: You can produce proteins that would normally kill a living host cell. Speed: It enables “benchtop” production in hours rather than days of cell culture. 2. Main Components Cell Extract (Lysate): The “machinery” (ribosomes, tRNAs, enzymes). DNA Template: The “instructions” for the protein. Energy System: ATP/GTP and a regeneration substrate (e.g., PEP). Amino Acids: The “building blocks.” Salts/Cofactors: Specifically $Mg^{2+}$ and $K^{+}$ for ribosome function. 3. Energy Provision Why it’s critical: Protein synthesis is energy-expensive. Without a regeneration system, ATP is depleted in minutes by side reactions, stopping production.Hugoen