Integration of cell-free protein synthesis and purification in one microfluidic chip for on-demand production of recombinant protein

Abstract

a) Xiaohai Yang and Kemin Wang conceived this project. Xiao Xiao designed and performed all the experiments. Yuan Zhou and Xiaobei Zhu fabricated the chip. Yuqiong Sun participated in polymerase chain reaction experiments. Qing Wang, Jianbo Liu, and Jin Huang provided helpful discussions. Xiao Xiao and Xiaohai Yang wrote the article.

Cell‐free protein synthesis: The transition from batch reactions to minimal cells and microfluidic devices

“…Accordingly, these compartments could proficiently provide a suitable home for CFPS reactions (see Figure 3). Additionally, artificial cells based on microbeads (Xiao et al, 2018), microgel (Jiao, Liu, Luo, Huck, & Yang, 2018), emulsions (Kang, Monsur Ali, Zhang, Pone, & Zhao, 2014), paper (Lim, Goh, & Khor, 2017), and vesicles (Elani, 2016) could be tailored for the generation of synthetic particles via microfluidic devices, which are further employed for the CFPS on the same chip (Damiati, Mhanna et al, 2018; Wang et al, 2018).…”

“…(c) Integrated cell‐free protein synthesis and purification in one microfluidic chip for on‐demand production of recombinant protein. Reprinted from Xiao et al (2018) with the permission of AIP Publishing. (d) Synthesis of minimal cells by incorporation of produced proteins by CFPS into lipid vesicles.…”

“…Being a mimetic porous vesicle, DNA hydrogels have been acknowledged as favorable platforms for functional and structural genomic studies when they are equipped with CFPS (Figure 6). Their porous structure allows for simple and swift diffusion of energy, biomolecules, and waste products, a key requirement for CFPS; meanwhile, they afford cell‐free gene/small interfering RNA (siRNA) synthesis and subsequent release of gene product on target cells (Xiao et al, 2018). In this sense, for gene therapy applications, in particular, stably expressing biomolecules within target cells is highly desirable, though yet an unrealizable effort.…”

“…on target cells (Xiao et al, 2018). In this sense, for gene therapy applications, in particular, stably expressing biomolecules within target cells is highly desirable, though yet an unrealizable effort.…”

The Usual Suspects 2019: of Chips, Droplets, Synthesis, and Artificial Cells

“…Since proteins for point-of-care applications require a certain purity, there remains a need to integrate protein synthesis and protein purification on a microfluidic chip in order to obtain the desired recombinant proteins with a simple operation. Xiao et al integrated two functional units, a protein synthesis unit and a protein purification unit, into a microfluidic chip for production of a recombinant protein (see Figure 4B) [58]. The first channel was filled with template DNA-modified agarose beads to form a cell-free protein synthesis unit, and the second channel was filled with nickel ion-modified agarose beads (Ni-nitrilotriacetic acid (NTA)) as a protein purification unit.…”