Expanding One-Pot Cell-Free Protein Synthesis and Immobilization for On-Demand Manufacturing of Biomaterials,ACS Synthetic Biology

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Expanding One-Pot Cell-Free Protein Synthesis and Immobilization for On-Demand Manufacturing of Biomaterials
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-02-23 00:00:00 , DOI: 10.1021/acssynbio.7b00383
Ana I. Benítez-Mateos ₁ , Irantzu Llarena ₂ , Ana Sánchez-Iglesias ₃ , Fernando López-Gallego _{1,

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Affiliation

Fabrication of protein-based biomaterials is an arduous and time-consuming procedure with multiple steps. In this work, we describe a portable toolkit that integrates both cell-free protein synthesis (CFPS) and protein immobilization in one pot just by mixing DNA, solid materials, and a CFPS system. We have constructed a modular set of plasmids that fuse the N-terminus of superfolded green fluorescent protein (sGFP) with different peptide tags (poly(6X)Cys, poly(6X)His, and poly(6X)Lys), which drive the immobilization of the protein on the tailored material (agarose beads with different functionalities, gold nanorods, and silica nanoparticles). This system also enables the incorporation of azide-based amino acids into the nascent protein for its selective immobilization through copper-free click reactions. Finally, this technology has been expanded to the synthesis and immobilization of enzymes and antibody-binding proteins for the fabrication of functional biomaterials. This synthetic biological platform has emerged as a versatile tool for on-demand fabrication of therapeutic, diagnostic, and sensing biomaterials.

中文翻译：

扩展用于生物材料按需生产的一锅无细胞蛋白质合成和固定化

蛋白质基生物材料的制造是一个繁琐且耗时的过程，需要多个步骤。在这项工作中，我们描述了一种便携式工具包，只需混合DNA，固体材料和CFPS系统，即可将无细胞蛋白质合成（CFPS）和蛋白质固定化整合到一个罐中。我们构建了一套模块化的质粒，该质粒将超折叠的绿色荧光蛋白（sGFP）的N端与不同的肽标签（poly（6X）Cys，poly（6X）His和poly（6X）Lys）融合在一起。将蛋白质固定在定制的材料上（具有不同功能的琼脂糖珠，金纳米棒和二氧化硅纳米颗粒）。该系统还能够将基于叠氮化物的氨基酸掺入到新生蛋白质中，以通过无铜点击反应将其选择性固定。最后，该技术已扩展到酶和抗体结合蛋白的合成和固定化，以制造功能性生物材料。这种合成生物平台已成为按需制造治疗，诊断和传感生物材料的多功能工具。

更新日期：2018-02-23

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