A microfluidic platform for sequential assembly and separation of synthetic cell models


Type
Article
Change log
Authors
Fletcher, Marcus 
Nahas, Kareem Al 
Abstract

Cell-sized vesicles like giant unilamellar vesicles (GUVs) are established as a promising biomimetic model for studying cellular phenomena in isolation. However, the presence of residual components and by-products, generated during vesicles preparation and manipulation, severely limits the utility of GUVs in applications like synthetic cells. Therefore, with the rapidly growing field of synthetic biology, there is an emergent demand for techniques that can continuously purify cell-like vesicles from diverse residues, while GUVs are being simultaneously synthesized and manipulated. We developed a microfluidic platform capable of purifying GUVs through stream bifurcation, where a stream of vesicles suspension is partitioned into three fractions - purified GUVs, residual components, and a washing solution. Using our purification approach, we showed that giant vesicles can be separated from various residues – that range in size and chemical composition – with a very high efficiency ( e = 0.99), based on size and deformability of the filtered objects. In addition, by incorporating the purification module with a microfluidic-based GUV-formation method, octanol-assisted liposome assembly (OLA), we established an integrated production-purification microfluidic unit that sequentially produces, manipulates, and purifies GUVs. We demonstrate the applicability of the integrated device to synthetic biology through sequentially fusing SUVs with freshly prepared GUVs and separating the fused GUVs from extraneous SUVs and oil droplets at the same time.

Description
Keywords
Journal Title
ACS Synthetic Biology
Conference Name
Journal ISSN
2161-5063
Volume Title
Publisher
American Chemical Society
Sponsorship
European Union Horizon 2020 Marie Sklodowska-Curie grant agreement No 892333, ERC consolidator grant (Designer- Pores 647144), EPSRC Doctoral Training Programme