Strategy exploration for developing robust lyophilized cell-free systems

Yang, Junzhu; Cui, Yuntao; Cao, Zhou; Ma, Shengnan; Lu, Yuan

doi:10.1016/j.biotno.2021.08.004

Cited by 15 publications

(19 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This enabled us to generate artificial cells within 30 min starting from the preparation of the lipid-oil to the formation of artificial cells. The successful protein synthesis using the lyophilized and rehydrated cell-free components has been reported previously ( Hunt et al, 2017 ; Lee et al, 2020 ; Yang et al, 2021 ). Therefore, it is possible to ship the samples at ambient temperature while avoiding moisture.…”

Section: Introductionmentioning

confidence: 87%

Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction

Shimane

Kuruma

2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Giant vesicles have been widely used for the bottom-up construction of artificial (or synthetic) cells and the physicochemical analysis of lipid membranes. Although methods for the formation of giant vesicles and the encapsulation of molecules within them have been established, a standardized protocol has not been shared among researchers including non-experts. Here we proposed a rapid and facile protocol that allows the formation of giant vesicles within 30 min. The quality of the giant vesicles encapsulating a cell-free protein expression system was comparable to that of the ones formed using a conventional method, in terms of the synthesis of both soluble and membrane proteins. We also performed protein synthesis in artificial cells using a lyophilized cell-free mixture and showed an equivalent level of protein synthesis. Our method could become a standard method for giant vesicle formation suited for artificial cell research.

show abstract

Section: Introductionmentioning

confidence: 87%

Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction

Shimane

Kuruma

2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…Two platforms are mainly involved in the development of riboswitch‐based biosensors, namely, the whole‐cell expression platform and the CFE platform (Z. Ma et al., 2020; J. Yang et al., 2021). Whole‐cell expression is created by engineering host cells to conditionally express fluorescent or colorimetric reporters only in the presence of the target of interest.…”

Section: Modular Design Of Riboswitch‐based Biosensormentioning

confidence: 99%

“…Two platforms are mainly involved in the development of riboswitch-based biosensors, namely, the whole-cell expression platform and the CFE platform (Z. Ma et al, 2020;J. Yang et al, 2021).…”

Section: Platformsmentioning

confidence: 99%

Advances in riboswitch‐based biosensor as food samples detection tool

Khan

Sun

et al. 2022

Comp Rev Food Sci Food Safe

View full text Add to dashboard Cite

Food safety has always been a hot issue of social concern, and biosensing has been widely used in the field of food safety detection. Compared with traditional aptamer-based biosensors, aptamer-based riboswitch biosensing represents higher precision and programmability. A riboswitch is an elegant example of controlling gene expression, where the target is coupled to the aptamer domain, resulting in a conformational change in the downstream expression domain and determining the signal output. Riboswitch-based biosensing can be extensively applied to the portable real-time detection of food samples. The numerous key features of riboswitch-based biosensing emphasize their sustainability, renewable, and testing, which promises to transform engineering applications in the field of food safety. This review covers recent developments in riboswitch-based biosensors. The brief history, definition, and modular design (regulatory mode, reporter, and expression platform) of riboswitch-based biosensors are explained for better insight into the design and construction. We summarize recent advances in various riboswitch-based biosensors involving theophylline, malachite green, tetracycline, neomycin, fluoride, thrombin, naringenin, ciprofloxacin, and paromomycin, aiming to provide general guidance for the design of riboswitch-based biosensors. Finally, the challenges and prospects are also summarized as a way forward stratagem and signs of progress.

show abstract

“…The Meningitis Vaccine Project recently benchmarked parameters for conjugate vaccine distribution in remote settings with the WHO approval of the MenAfriVac vaccine for controlled temperature chain storage for 4 days at up to 40 °C with a cost of <$0.50 per dose. , Adjusting CFE reaction formulations could address these challenges in our cell-free conjugate vaccine production platform. However, to date, CFE optimizations have typically sought to address either cost − or thermostability − rather than considering both formulation properties together.…”

Section: Introductionmentioning

confidence: 99%

A Low-Cost, Thermostable, Cell-Free Protein Synthesis Platform for On-Demand Production of Conjugate Vaccines

et al. 2022

View full text Add to dashboard Cite

Cell-free protein synthesis systems that can be lyophilized for long-term, non-refrigerated storage and transportation have the potential to enable decentralized biomanufacturing. However, increased thermostability and decreased reaction cost are necessary for further technology adoption. Here, we identify maltodextrin as an additive to cell-free reactions that can act as both a lyoprotectant to increase thermostability and a low-cost energy substrate. As a model, we apply optimized formulations to produce conjugate vaccines for ∼$0.50 per dose after storage at room temperature (∼22 °C) or 37 °C for up to 4 weeks, and ∼$1.00 per dose after storage at 50 °C for up to 4 weeks, with costs based on raw materials purchased at the laboratory scale. We show that these conjugate vaccines generate bactericidal antibodies against enterotoxigenic Escherichia coli (ETEC) O78 O-polysaccharide, a pathogen responsible for diarrheal disease, in immunized mice. We anticipate that our low-cost, thermostable cell-free glycoprotein synthesis system will enable new models of medicine biosynthesis and distribution that bypass cold-chain requirements.

show abstract

Strategy exploration for developing robust lyophilized cell-free systems

Cited by 15 publications

References 31 publications

Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction

Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction

Advances in riboswitch‐based biosensor as food samples detection tool

A Low-Cost, Thermostable, Cell-Free Protein Synthesis Platform for On-Demand Production of Conjugate Vaccines

Contact Info

Product

Resources

About