Photothermal heating of cell-free reactions for on-site production of recombinant proteins

Cell-free synthesis technology is emerging as a versatile platform for biomanufacturing, particularly for on-site production of essential products. In this study, we address a pivotal challenge encountered when utilizing cell-free protein synthesis (CFPS) for on-site production in resource-limited settings. The efficacy of CFPS critically depends on precise temperature regulation. However, traditional heating methods are impractical for application outside of laboratory environments. To address this challenge, we propose an innovative solution that integrates gold nanorods (GNRs) with photothermal properties into the CFPS reaction mixture. Upon activation by a handheld laser module emitting near-infrared light, these GNRs efficiently convert light energy into heat, enabling rapid and precise temperature control for protein synthesis. Our approach not only achieves optimal synthesis temperatures more rapidly than conventional methods but also significantly enhances protein yields, with an increase of over threefold within a 30-min reaction period. In summary, our findings highlight the potential of photothermal heating as a means to ensure portable and efficient protein synthesis in the field. This advancement brings CFPS closer to achieving real-time, on-site protein production.