Microfluidic platform for rapid screening of bacterial cell lysis

Fradique, Ricardo; Azevedo, Ana M.; Chu, V.; Conde, J.P.; Aires-Barros, M. Raquel

doi:10.1016/j.chroma.2019.460539

Cited by 8 publications

(18 citation statements)

References 36 publications

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“…It was shown that even a significant increase in the concentration of the lysis reagent does not increase the lysis efficiency of the cell solution. On the other hand, increasing the volume fraction of the lysis reagent (the ratio of the volume initially filled with the lysis reagent to the total volume of the droplet) to 97%, achieved a complete cell solution lysis within 0.25 s. R. Fradique et al reported a diffusion-based microfluidic device for the rapid screening of continuous lysis conditions, utilizing E. coli as a model host [ 18 ]. The main goal of the work was to determine the efficiency of different lysis conditions for the release of an intracellular protein from the bacteria cells, in other words, to measure the effect of different lysis solutions, contact times, and cell to lysis solution volume ratios.…”

Section: Cell Lysis Methodsmentioning

confidence: 99%

“…diffusion-based microfluidic device for the rapid screening of continuous lysis conditions, utilizing E. coli as a model host [18]. The main goal of the work was to determine the efficiency of different lysis conditions for the release of an intracellular protein from the bacteria cells, in other words, to measure the effect of different lysis solutions, contact times, and cell to lysis solution volume ratios.…”

Section: Chemical Lysismentioning

confidence: 99%

“…The authors utilized a silica porous polymer monolith in the presence of a lysis agent (sodium dodecyl sulfate (SDS) and TritonX-100 at different concentrations), to produce additional shear and friction forces to the pressure-driven solution flow [21]. The lysis was confirmed by [18] for buffer resuspended cells, using Tris-HCl buffer, BPER, and lysozyme. (a) For different total flow rates (using the same flow rates of resuspended cell solution and buffer or lysis solution); (b) for different flow rate ratios (keeping constant total flow rate).…”

Section: Chemical Lysismentioning

confidence: 99%

“…( a ) For different total flow rates (using the same flow rates of resuspended cell solution and buffer or lysis solution); ( b ) for different flow rate ratios (keeping constant total flow rate). Adapted with permission from [ 18 ]. Copyright 2020 Copyright Elsevier.…”

Section: Figurementioning

confidence: 99%

“…Figure 4. Percentage of cell lysis obtained in[18] for buffer resuspended cells, using Tris-HCl buffer, BPER, and lysozyme. (a) For different total flow rates (using the same flow rates of resuspended cell solution and buffer or lysis solution); (b) for different flow rate ratios (keeping constant total flow rate).…”

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confidence: 99%

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Review of Microfluidic Methods for Cellular Lysis

et al. 2021

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Cell lysis is a process in which the outer cell membrane is broken to release intracellular constituents in a way that important information about the DNA or RNA of an organism can be obtained. This article is a thorough review of reported methods for the achievement of effective cellular boundaries disintegration, together with their technological peculiarities and instrumental requirements. The different approaches are summarized in six categories: chemical, mechanical, electrical methods, thermal, laser, and other lysis methods. Based on the results derived from each of the investigated reports, we outline the advantages and disadvantages of those techniques. Although the choice of a suitable method is highly dependent on the particular requirements of the specific scientific problem, we conclude with a concise table where the benefits of every approach are compared, based on criteria such as cost, efficiency, and difficulty.

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Section: Cell Lysis Methodsmentioning

confidence: 99%

Section: Chemical Lysismentioning

confidence: 99%

Section: Chemical Lysismentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

mentioning

confidence: 99%

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Review of Microfluidic Methods for Cellular Lysis

et al. 2021

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Continuous microfluidic platform combining cell lysis and protein extraction for screening overall process conditions

Fradique,

Jardim,

Azevedo

et al. 2023

J of Chemical Tech & Biotech

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BACKGROUNDMicrofluidic devices have been increasingly used as tools to accelerate the development of new biomanufacturing methods, given their ability to test a large set of variables in a short amount of time with minimal reagent consumption. However, while being able to expedite a bioprocess design, the modular aspect of these devices has not been systematically explored, with most applications focusing on single unitary operations. This paper explores the modular design of these devices through the development of an integrated microfluidic chip.RESULTSTwo separate unitary operations were included into a single device for continuous operation: product release by chemical cell lysis and product concentration by aqueous two‐phase system. In this way, it is possible to screen multiple conditions for each operation and evaluate their combined effect on the final product, making this a great tool in the development and optimization of a process. A recombinant Escherichia coli (E. coli) strain producing green fluorescent protein (GFP) was used as model system, which allows for lysis efficiency and partition coefficient to be evaluated by fluorescence microscopy. A chemical lysis solutions was used in the lysis step, presenting lysis efficiencies of about 100%, while a polyethylene glycol (PEG)/phosphate system was screened for the separation stage, presenting partition coefficients of about 4.CONCLUSIONSThe integrated device allowed for the continuous screening of multiple combinations of operation conditions, highlighting, for example, conditions where improvements on the lysis efficiency subsequently impaired the separation, decreasing the partition of the target product, thus demonstrating the need for the evaluation of a process in its entirety. In this way, the microfluidic device delivered a rapid process screening and optimization, with very low reagent consumption, and using a simple fluorescence microscope for data collection. © 2023 Society of Chemical Industry (SCI).

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