Cryopreservation of a cell-based biosensor chip modified with elastic polymer fibers enabling ready-to-use on-site applications

Özsoylu, Dua; Işık, Tuğba; Demir, Mustafa M.; Schöning, Michael J.; Wagner, Torsten

doi:10.1016/j.bios.2021.112983

Cited by 20 publications

(11 citation statements)

References 54 publications

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“…To this end, we tested several selected cryoprotectants that could provide extracellular and intracellular protection to the cells during storage under freezing conditions [24] to enhance the long-term storability and maintain the quantitative determination capability of the array for target amino acids. Although several studies have reported the benefits of cell cryopreservation [23,25], the cryopreservation of microbial cell-based amino acid arrays has not been previously reported. Six cryoprotectants (DMSO, glycerol, EG, PEG, sucrose, and trehalose), which have been widely utilized as cryoprotectants for preserving the cell viability, were added to individual wells of a 96-well plate containing Met auxotroph cultures in agarose; the mixture was then allowed to solidify at room temperature.…”

Section: Resultsmentioning

confidence: 99%

“…To overcome the limitations of delivering each component to the end users separately, the entire amino acid array system can be prepared at the manufacturing stage and transported to the end users in a ready-to-use format [22]. However, in this case, the cells within the array should be carefully preserved to maintain their viability during storage and transportation [23]. Therefore, the storability of the cells is critical for the practical applications of the amino acid array platform.…”

Section: Introductionmentioning

confidence: 99%

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Effective Cryopreservation of a Bioluminescent Auxotrophic Escherichia coli-Based Amino Acid Array to Enable Long-Term Ready-to-Use Applications

et al. 2021

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Amino acid arrays comprising bioluminescent amino acid auxotrophic Escherichia coli are effective systems to quantitatively determine multiple amino acids. However, there is a need to develop a method for convenient long-term preservation of the array to enable its practical applications. Here, we reported a potential strategy to efficiently maintain cell viability within the portable array. The method involves immobilization of cells within agarose gel supplemented with an appropriate cryoprotectant in individual wells of a 96-well plate, followed by storage under freezing conditions. Six cryoprotectants, namely dimethyl sulfoxide, glycerol, ethylene glycol, polyethylene glycol, sucrose, and trehalose, were tested in the methionine (Met) auxotroph-based array. Carbohydrate-type cryoprotectants (glycerol, sucrose, and trehalose) efficiently preserved the linearity of determination of Met concentration. In particular, the array with 5% trehalose exhibited the best performance. The Met array with 5% trehalose could determine Met concentration with high linearity (R2 value = approximately 0.99) even after storage at −20 °C for up to 3 months. The clinical utilities of the Met and Leu array, preserved at −20 °C for 3 months, were also verified by successfully quantifying Met and Leu in spiked blood serum samples for the diagnosis of the corresponding metabolic diseases. This long-term preservation protocol enables the development of a ready-to-use bioluminescent E. coli-based amino acid array to quantify multiple amino acids and can replace the currently used laborious analytical methods.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effective Cryopreservation of a Bioluminescent Auxotrophic Escherichia coli-Based Amino Acid Array to Enable Long-Term Ready-to-Use Applications

et al. 2021

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“…This device enhanced, controlled, and offered a continuous CPA loading without damaging the spherical morphology of blastocytes and minimized the shrinkage rate. Moreover, in a recent study, Özsoylu et al developed a method to preserve the cell-based biosensor chip, which allowed one to preserve adherent cells on sensor surfaces and, therefore, this method was called on-sensor cryopreservation (OSC) [ 482 ]. Here, the biosensor surface was modified by polyethylene vinyl acetate (PEVA) electrospun fibers to make the sensor durable for high temperature changes.…”

Section: Biomedical Applicationsmentioning

confidence: 99%

Biomedical Applications of Microfluidic Devices: A Review

et al. 2022

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Both passive and active microfluidic chips are used in many biomedical and chemical applications to support fluid mixing, particle manipulations, and signal detection. Passive microfluidic devices are geometry-dependent, and their uses are rather limited. Active microfluidic devices include sensors or detectors that transduce chemical, biological, and physical changes into electrical or optical signals. Also, they are transduction devices that detect biological and chemical changes in biomedical applications, and they are highly versatile microfluidic tools for disease diagnosis and organ modeling. This review provides a comprehensive overview of the significant advances that have been made in the development of microfluidics devices. We will discuss the function of microfluidic devices as micromixers or as sorters of cells and substances (e.g., microfiltration, flow or displacement, and trapping). Microfluidic devices are fabricated using a range of techniques, including molding, etching, three-dimensional printing, and nanofabrication. Their broad utility lies in the detection of diagnostic biomarkers and organ-on-chip approaches that permit disease modeling in cancer, as well as uses in neurological, cardiovascular, hepatic, and pulmonary diseases. Biosensor applications allow for point-of-care testing, using assays based on enzymes, nanozymes, antibodies, or nucleic acids (DNA or RNA). An anticipated development in the field includes the optimization of techniques for the fabrication of microfluidic devices using biocompatible materials. These developments will increase biomedical versatility, reduce diagnostic costs, and accelerate diagnosis time of microfluidics technology.

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“…LAPS can be designed as multiwell-and multianalyte-sensor devices and can serve for chemical imaging, where the distribution of the analyte concentration is visualized on its chip surface [10][11][12][13][14]. Moreover, LAPS provides a broad range of possible applications and has been utilized for various (bio)chemical and biotechnological approaches, such as monitoring the metabolic activity of bacteria in fermentation broth [14], for on-sensor cryopreservation of cells [15], multi-ion and penicillin detection [16,17], and DNA sensing [18].…”

Section: Introductionmentioning

confidence: 99%

Light-Addressable Actuator-Sensor Platform for Monitoring and Manipulation of pH Gradients in Microfluidics: A Case Study with the Enzyme Penicillinase

et al. 2021

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The feasibility of light-addressed detection and manipulation of pH gradients inside an electrochemical microfluidic cell was studied. Local pH changes, induced by a light-addressable electrode (LAE), were detected using a light-addressable potentiometric sensor (LAPS) with different measurement modes representing an actuator-sensor system. Biosensor functionality was examined depending on locally induced pH gradients with the help of the model enzyme penicillinase, which had been immobilized in the microfluidic channel. The surface morphology of the LAE and enzyme-functionalized LAPS was studied by scanning electron microscopy. Furthermore, the penicillin sensitivity of the LAPS inside the microfluidic channel was determined with regard to the analyte’s pH influence on the enzymatic reaction rate. In a final experiment, the LAE-controlled pH inhibition of the enzyme activity was monitored by the LAPS.

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Cryopreservation of a cell-based biosensor chip modified with elastic polymer fibers enabling ready-to-use on-site applications

Cited by 20 publications

References 54 publications

Effective Cryopreservation of a Bioluminescent Auxotrophic Escherichia coli-Based Amino Acid Array to Enable Long-Term Ready-to-Use Applications

Effective Cryopreservation of a Bioluminescent Auxotrophic Escherichia coli-Based Amino Acid Array to Enable Long-Term Ready-to-Use Applications

Biomedical Applications of Microfluidic Devices: A Review

Light-Addressable Actuator-Sensor Platform for Monitoring and Manipulation of pH Gradients in Microfluidics: A Case Study with the Enzyme Penicillinase

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