Biotechnology and Bioprocess Engineering 2024; 29(2): 255-261  
Photothermal heating of cell‑free reactions for on‑site production of recombinant proteins
Kyunghwan Yeom1 · Yu Jin Park2 · Hansol Kim1 · Dong‑Yeon Song2 · Dong‑Myung Kim2 · Ji‑Ho Park1,3
1 Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea
2 Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon 34134, Korea
3 KAIST Institute for Health Science and Technology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea
Correspondence to: Dong‑Myung Kim
dmkim@cnu.ac.kr
Ji‑Ho Park
jihopark@kaist.ac.kr

Kyunghwan Yeom and Yu Jin Park have contributed equally to this work.
Received: October 27, 2023; Revised: November 13, 2023; Accepted: November 16, 2023; Published online: March 14, 2024.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
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.
Keywords: Cell-free protein synthesis · Gold nanorods · Photothermal effect · On-demand biomanufacturing

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