2025/12/25 by Byeongmin Chae, Hyeongwoo Park, Hyewon Jeon +6 · 1 voice
Biochemistry, Genetics and Molecular Biology · Medicine · #RNA and protein synthesis mechanisms #Monoclonal and Polyclonal Antibodies Research #Viral Infectious Diseases and Gene Expression in Insects
paper · pdf · doi:10.64898/2025.12.23.696326
openalex publication_date 2025/12/25 · openalex created_date 2025/12/26 · openalex updated_date 2026/07/31
High-performance cell-free protein synthesis has transformative potential for synthetic biology, yet the prohibitive costs of PURE kits and the labor intensity of in-house preparation have restricted accessibility and scalability. We developed i-POPFLEX (Purified components OPtimized for FLEXible protein expression), a modular cell-free protein synthesis system in which 34 translation factors, individually synthesized in vitro, are assembled using automated liquid handling. This workflow minimizes manual input and supports parallelized production, generating complete, ready-to-use systems within two days. i-POPFLEX achieves up to 5-fold higher protein yields and a 95 % cost reduction (20-fold lower cost) compared with commercial kits. Its flexible architecture also enables selective component inclusion for genetic code reprogramming and site-specific incorporation of non-canonical amino acids. By coupling modular design with automation, i-POPFLEX provides an accessible, customizable, and economically viable platform for next-generation biomanufacturing workflows.