Microcompartmentalized Cell-Free Protein Synthesis in Hydrogel μ-Channels,ACS Synthetic Biology

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Microcompartmentalized Cell-Free Protein Synthesis in Hydrogel μ-Channels
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-11-10 , DOI: 10.1021/acssynbio.0c00462
Ana I Benítez-Mateos _{1,

2,

3} , Nicoll Zeballos ₁ , Natalia Comino ₁ , Lucía Moreno de Redrojo ₂ , Teodora Randelovic _{4,

5} , Fernando López-Gallego _{1,

2,

6,

7}

Affiliation

The rapid demand for protein-based molecules has stimulated much research on cell-free protein synthesis (CFPS); however, there are still many challenges in terms of cost-efficiency, process intensification, and sustainability. Herein, we describe the microcompartmentalization of CFPS of superfolded green fluorescent protein (sGFP) in alginate hydrogels, which were casted into a μ-channel device. CFPS was optimized for the microcompartmentalized environment and characterized in terms of synthesis yield. To extend the scope of this technology, the use of other biocompatible materials (collagen, laponite, and agarose) was explored. In addition, the diffusion of sGFP from the hydrogel microenvironment to the bulk was demonstrated, opening a promising opportunity for concurrent synthesis and delivery of proteins. Finally, we provide an application for this system: the CFPS of enzymes. The present design of the hydrogel μ-channel device may enhance the potential application of microcompartmentalized CFPS in biosensing, bioprototyping, and therapeutic development.

中文翻译：

水凝胶 μ 通道中的微区室化无细胞蛋白质合成

对基于蛋白质的分子的快速需求刺激了对无细胞蛋白质合成 (CFPS) 的大量研究。然而，在成本效率、流程集约化和可持续性方面仍然存在许多挑战。在此，我们描述了海藻酸盐水凝胶中超折叠绿色荧光蛋白 (sGFP) 的 CFPS 的微区室化，这些水凝胶被浇铸到 μ 通道装置中。CFPS 针对微区室环境进行了优化，并在合成产率方面进行了表征。为了扩展这项技术的范围，探索了使用其他生物相容性材料（胶原蛋白、合成锂皂石和琼脂糖）。此外，还证明了 sGFP 从水凝胶微环境向整体的扩散，为同时合成和递送蛋白质开辟了广阔的前景。最后，我们为这个系统提供了一个应用：酶的 CFPS。水凝胶 μ 通道装置的当前设计可能会增强微室化 CFPS 在生物传感、生物原型设计和治疗开发中的潜在应用。

更新日期：2020-11-21

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