Dataset Information

Cell-Free Synthesis Goes Electric: Dual Optical and Electronic Biosensor via Direct Channel Integration into a Supported Membrane Electrode.

ABSTRACT: Assembling transmembrane proteins on organic electronic materials is one promising approach to couple biological functions to electrical readouts. A biosensing device produced in such a way would enable both the monitoring and regulation of physiological processes and the development of new analytical tools to identify drug targets and new protein functionalities. While transmembrane proteins can be interfaced with bioelectronics through supported lipid bilayers (SLBs), incorporating functional and oriented transmembrane proteins into these structures remains challenging. Here, we demonstrate that cell-free expression systems allow for the one-step integration of an ion channel into SLBs assembled on an organic conducting polymer, poly(3,4-ethylenedioxythiophene) polystyrenesulfonate (PEDOT:PSS). Using the large conductance mechanosensitive channel (MscL) as a model ion channel, we demonstrate that MscL adopts the correct orientation, remains mobile in the SLB, and is active on the polyelectrolyte surface using optical and electrical readouts. This work serves as an important illustration of a rapidly assembled bioelectronic platform with a diverse array of downstream applications, including electrochemical sensing, physiological regulation, and screening of transmembrane protein modulators.

SUBMITTER: Manzer ZA

PROVIDER: S-EPMC11364431 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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Cell-Free Synthesis Goes Electric: Dual Optical and Electronic Biosensor via Direct Channel Integration into a Supported Membrane Electrode.

Manzer Zachary A ZA Ghosh Surajit S Roy Arpita A Jacobs Miranda L ML Carten Juliana J Kamat Neha P NP Daniel Susan S

ACS synthetic biology 20230118 2

Assembling transmembrane proteins on organic electronic materials is one promising approach to couple biological functions to electrical readouts. A biosensing device produced in such a way would enable both the monitoring and regulation of physiological processes and the development of new analytical tools to identify drug targets and new protein functionalities. While transmembrane proteins can be interfaced with bioelectronics through supported lipid bilayers (SLBs), incorporating functional ...[more]

PMID: 36651574

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Project description:Vaping (inhalation of electronic cigarette-generated aerosol) is a public health concern. Due to recent spikes in adolescent use of electronic cigarettes (ECIGs) and vaping-induced illnesses, demand for scientific inquiry into the physiological effects of electronic cigarette (ECIG) aerosol has increased. For such studies, standardized and consistent aerosol production is required. Many labs generate aerosol by manually activating peristaltic pumps and ECIG devices simultaneously in a predefined manner. The tedium involved with this process (large puff number over time) and risk of error in keeping with puff topography (puff number, duration, interval) are less than optimal. Furthermore, excess puffing on an ECIG device results in battery depletion, reducing aerosol production, and ultimately, its chemical and physical nature. While commercial vaping machines are available, the cost of these machines is prohibitive to many labs. For these reasons, an economical and programmable ECIG aerosol generator, capable of generating aerosol from two atomizers simultaneously, was fabricated, and subsequently validated. Validation determinants include measurements of atomizer temperatures (inside and outside), electrical parameters (current, resistance and power) of the circuitry, aerosol particle distribution (particle counts and mass concentrations) and aerosol delivery (indexed by nicotine recovery), all during stressed conditions of four puffs/minute for 75 min (i.e., 300 puffs). Validation results indicate that the ECIG aerosol generator is better suited for experiments involving ≤100 puffs. Over 100 puffs, the amount of variation in the parameters measured tends to increase. Variations between channels are generally higher than variations within a channel. Despite significant variations in temperatures, electrical parameters, and aerosol particle distributions, both within and between channels, aerosol delivery remains remarkably stable for up to 300 puffs, yielding over 25% nicotine recovery for both channels. In conclusion, this programmable, dual-channel ECIG aerosol generator is not only affordable, but also allows the user to control puff topography and eliminate battery drain of ECIG devices. Consequently, this aerosol generator is valid, reliable, economical, capable of using a variety of E-liquids and amenable for use in a vast number of studies investigating the effects of ECIG-generated aerosol while utilizing a multitude of puffing regimens in a standardized manner.

Dataset Information

Cell-Free Synthesis Goes Electric: Dual Optical and Electronic Biosensor via Direct Channel Integration into a Supported Membrane Electrode.

Publications

Cell-Free Synthesis Goes Electric: Dual Optical and Electronic Biosensor via Direct Channel Integration into a Supported Membrane Electrode.

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