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. 2018 Aug 10;8:11967. doi: 10.1038/s41598-018-30435-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Removal of HeLa cell lysate based CFPS media from the solid support after expression and capture. (A) Split-intein mediated capture (split-intein mediated capture) after CFPS in reaction matrix before immobilization, followed by immobilization, washing, and light triggered release (light triggered release). Lanes: (1) molecular weight marker; (2) Peptide 3; (3) HeLa lysate without DNA (negative control); (4) hCNTF-NpuDnaE_ΔC16 CFPS expression without TCEP; (5) hCNTF-NpuDnaE_ΔC16 CFPS expression with 1 mM TCEP; (6) hCNTF- NpuDnaE_ΔC16 after protein trans splicing with peptide 3; (7–9) wash steps 1, 2 and 3; (10–15) 0, 15, 30, 60, 180, and 360 minutes of light triggered release (365 nm, 12.5 mW). (B) Peptide immobilization followed by split-intein mediated capture, washing, and light triggered release. Lanes: (1) molecular weight marker; (2) HeLa lysate without DNA (negative control); (3) hCNTF-NpuDnaE_ΔC16 CFPS expression without TCEP; (4) hCNTF- NpuDnaE_ΔC16 CFPS expression after protein trans splicing with peptide 3; (5–7) wash steps 1, 2 and 3; (8–13) 0, 15, 30, 60, 180, and 360 minutes of light triggered release (365 nm, 12.5 mW). Uncropped images can be seen in Fig. S8.