Electrochemical H₂O₂ biosensor composed of myoglobin on MoS₂ nanoparticle-graphene oxide hybrid structure

Yoon, Jinho,Lee, Taek,Bapurao G., Bharate,Jo, Jinhee,Oh, Byung-Keun,Choi, Jeong-Woo Elsevier 2017 Biosensors & bioelectronics Vol.93 No.-

<P><B>Abstract</B></P> <P>In this research, the electrochemical biosensor composed of myoglobin (Mb) on molybdenum disulfide nanoparticles (MoS<SUB>2</SUB> NP) encapsulated with graphene oxide (GO) was fabricated for the detection of hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>). Hybrid structure composed of MoS<SUB>2</SUB> NP and GO (GO@MoS<SUB>2</SUB>) was fabricated for the first time to enhance the electrochemical signal of the biosensor. As a sensing material, Mb was introduced to fabricate the biosensor for H<SUB>2</SUB>O<SUB>2</SUB> detection. Formation and immobilization of GO@MoS<SUB>2</SUB> was confirmed by transmission electron microscopy, ultraviolet-visible spectroscopy, scanning electron microscopy, and scanning tunneling microscopy. Immobilization of Mb, and electrochemical property of biosensor were investigated by cyclic voltammetry and amperometric i-t measurements. Fabricated biosensor showed the electrochemical signal enhanced redox current as −1.86μA at an oxidation potential and 1.95μA at a reduction potential that were enhanced relative to those of electrode prepared without GO@MoS<SUB>2</SUB>. Also, this biosensor showed the reproducibility of electrochemical signal, and retained the property until 9 days from fabrication. Upon addition of H<SUB>2</SUB>O<SUB>2</SUB>, the biosensor showed enhanced amperometric response current with selectivity relative to that of the biosensor prepared without GO@MoS<SUB>2</SUB>. This novel hybrid material-based biosensor can suggest a milestone in the development of a highly sensitive detecting platform for biosensor fabrication with highly sensitive detection of target molecules other than H<SUB>2</SUB>O<SUB>2</SUB>.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The electrochemical biosensor composed of the myoglobin on the graphene oxide (GO)-encapsulated molybdenum disulfide nanoparticles (MoS<SUB>2</SUB> NP) was developed for hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>) detection. </LI> <LI> MoS<SUB>2</SUB> NP were encapsulated by GO (GO@MoS<SUB>2</SUB>) for the first time to induce the electrochemical signal enhancement. </LI> <LI> Fabricated electrochemical biosensor with GO@MoS<SUB>2</SUB> showed the stability and reproducibility. </LI> <LI> Fabricated electrochemical biosensor also showed the enhanced amperometric response current by detection of H<SUB>2</SUB>O<SUB>2</SUB>. </LI> </UL> </P>

Comparison and evaluation of the performance of graphene-based biosensors

Abdelbasset Walid Kamal,Jasim Saade Abdalkareem,Bokov Dmitry Olegovich,Oleneva Maria Sergeevna,Islamov Anvar,Hammid Ali Thaeer,Mustafa Yasser Fakri,Yasin Ghulam,Alguno Arnold C.,Kianfar Ehsan 한국탄소학회 2022 Carbon Letters Vol.32 No.4

Biosensors are a group of measurement systems and their design is based on the selective identification of analyses based on biological components and physical and chemical detectors. Biosensors consist of three components: biological element, detector, and converter. The design of biosensors in various fields of biological sciences, medicine has expanded significantly. Biosensor technology actually represents a combination of biochemistry, molecular biology, chemistry, physics, electronics, and telecommunications. A biosensor actually consists of a small sensor and biological material fixed on it. Because biosensors are a powerful tool for identifying biological molecules, today they are used in various medical sciences, chemical industry, food industry, environmental monitoring, pharmaceutical production, health, etc. In fact, these sensors are a powerful tool to identify biological molecules. In fact, biosensors are analytical tools that can use biological intelligence to detect and react with a compound or compounds, and thus create a chemical, optical, or electrical message. The basis of a biosensor is to convert a biological response into a message. In this category, the use of telecommunication engineering technology and electromagnetic waves and frequency and radio spectrum is growing more and more to detect, measure, and determine the desired parameters in microbiology and laboratory sciences. The use of radio, optical, electromagnetic, ultrasonic, and infrared wave detection technology is part of the applications of telecommunication science in this field. Even image and audio processing systems have been instrumental in the discussion of biosensors in microbiology. The science of using fiber optics and waveguides, micro-strip antennas, and microelectromechanical technology is also very efficient in the construction and design of these biosensors.

Recent advances in the metamaterial-inspired biosensors

Salim, Ahmed,Lim, Sungjoon Elsevier 2018 Biosensors & bioelectronics Vol.117 No.-

<P><B>Abstract</B></P> <P>Metamaterials (MM)-inspired microwave biosensors are a valuable addition to the field of diagnostic approaches and prognostic tools. The fundamental principle behind these biosensors is unique dielectric signatures corresponding to healthy/diseased tissues. Relying on nonionizing radiation and offering an increased resolution with accuracy comparable to that of ultrasound devices, they are an attractive solution for noninvasive and label-free biosensing applications. High-quality-factor MM-inspired resonators are integrated with microfluidics to accelerate the lab-on-chip and point-of-care diagnostic approaches owing to the small detection volume and overall compact size of these devices. A variety of biomolecular detection, glucose detection and hyperthermia treatment using state-of-the-art MM-inspired biosensors have been discussed. Optical transduction techniques (e.g., surface plasmon resonance) which enhance the sensitivity in terms of limit-of-detection and resolution, have also been outlined. Utilization of microwave biosensors as therapeutic agents is at its initial stages owing to lack of required sensitivity and reliability in recently proposed MM-inspired biosensors.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Commonly used materials and fabrication techniques for RF biosensors have been studied. </LI> <LI> Design strategies and simulation setup of RF biosensors have been studied. </LI> <LI> The most recent metamaterial-inspired biosensors have been reviewed by dividing GHz and THz domains. </LI> <LI> Finally, challenges and resolving strategies for metamaterial-inspired biosensors are discussed. </LI> </UL> </P>

FACS-seq as a Powerful Tool for Profiling the Dose-response Curves of Biosensors in a Massively Parallel Manner

Chong ZHANG 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.4

Living organisms have a variety of mechanisms to sense and respond to the environmental signals by dynamically regulating their genetic expression networks. Harnessing this ability, genetically encoded biosensors, mimicking natural regulation networks by assembling basic biological parts like promoter, ribosome binding site, operator, reporter etc. into genetic circuit, are developed to recognize the analytes and transduce their signals to a measurable output. Dose-response curve, mapping the genetic circuits to their function, shapes how individual biosensors respond to the specific signals, which is crucial not only for the specific usage scenario of genetic encoded biosensor, but also for illustrating their regulatory functions in living cells. In this report, we would like show our recent attempts on the application of a dose-response profiling method, namely fluorescence-activated cell sorting coupled with next-generation sequencing (FACS-seq), in generating accurate dose-response curves for thousands of biosensors in a massively parallel manner, which provides a powerful platform for dissecting the mechanistic basis of the regulatory elements in living cells, and for the fine tuning of biosensors in a customized and low-cost manner. As the first example, we focused on tnaC, which encodes the tryptophan operon leader peptide in bacteria and is a model of macromolecular-machinery-dynamics-dependent regulatory elements. Working as a molecular sensor, tnaC responds to intracellular tryptophan (Trp) and regulates the biosynthesis of indol. We used FACS-seq to generate accurate response curves for all possible codon substitutions in tnaC. The FACS-seq results allowed us to generate comprehensive genotype-phenotype maps for 1,450 tnaC variants in living cells. The results clarified the nature of several transient, previously enigmatic states involving RNAP and the ribosome, and these states play important roles in the tnaC sensor response. Using in silico modeling and additional experiment, we further demonstrated the molecular basis of the quantitative relation between basal and inductive response, as well as the range of detection of the sensor. In the second example, we developed a novel biological parts assembly workflow to encode genetic circuits with short DNA barcodes, which make sure one-to-one correspondence of the barcodes and biological parts combinations, enabling high-throughput generation of dose-response curves of higher-order combinatorial biosensor space. As a proof of concept, we applied our novel workflow for the fine-tuning of the dose-response curve of malonyl-CoA biosensor based on FapR-fapO system in Saccharomyces cerevisiae. With our 5-step encoding workflow, a trackable combinatorial library contained 5155/5184 combinations with 6 levels of TF dosage, 4 different operator positions, and 216 possible UAS designs, was constructed. FACS-seq successfully characterized the response curve of 2,632 biosensors out of 5184 combinations, providing large-scale genotype-phenotype association data of the designed biosensors. Machine-learning algorithms were then developed to predict uncharacterized dose-response curves and identify key features in the whole combinatorial library, generating a panoramic scanning map of the combinatorial space. With the assistance of our novel workflow, 3755 dose-response curves were obtained at a cost of $1.37 per curve, and a malonyl-CoA biosensor with the largest dynamic response range reported so far was successfully acquired.

Reusable and storable whole-cell microbial biosensors with a microchemostat platform for in situ on-demand heavy metal detection

Bae, Juyeol,Lim, Ji-Won,Kim, Taesung Elsevier 2018 Sensors and actuators. B Chemical Vol.264 No.-

<P><B>Abstract</B></P> <P>Target analyte detection using whole-cell microbial biosensors can benefit from a microchemostat platform owing to the reduced reagent consumption, a finely controllable environment, and multi-parallel processing. However, the use of the previous microchemostat still has limitations for practical applications, e.g., its single-use design, long time requirements for cell and device preparation, low portability, slow and weak response signals, and the lack of ready-to-use in situ devices. In the present study, we developed a novel microchemostat platform that resolves these issues and enhances the performance of microbial biosensors via its abilities to actively control the cell population and to form a uniform culture environment using a nanoscale hydrodynamic film (NHF). The combination of the microchemostat platform and the microbial biosensors yielded fast and strong signals on demand in response to heavy metal ions (e.g., Pb<SUP>2+</SUP>). In addition, the platform enables the long-term on-chip storage of microbial biosensors for over 1 month, without deterioration of growth and detection ability. Lastly, we demonstrated that microbial biosensors can consecutively measure multiple samples by a regeneration process in which the microchemostat platform continuously subcultures the microbial biosensors until the complete recovery of their sensing capabilities. Thus, the microchemostat offers whole-cell microbial biosensors that are fast, ready-to-use, reusable, and storable. It also shows good potential for user-friendly, portable on-site environmental monitoring, particularly where expensive analytical tools and/or instruments for heavy metal ion detection are unavailable.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Our microchemostat platform addresses most practical limitations of existing platforms for whole-cell microbial biosensors. </LI> <LI> Long-term on-chip storage of cells under refrigeration can relieve end-users from cumbersome off-chip cell preparation. </LI> <LI> The microchemostat can regenerate used biosensors for multiple analyses with only a single chip. </LI> <LI> The microchemostat platform shows remarkable potential for on-site, in situ, and on-demand heavy metal ions detection. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>The microchemostat platform can make the whole-cell microbial biosensors reusable, ready-to-use, and storable, showing a dramatic potential for portable, on-site, in situ, and on‐demand environmental monitoring of heavy metal ions.</P> <P>[DISPLAY OMISSION]</P>

Genetically Encoded Biosensor Engineering for Application in Directed Evolution

Mao Yin,Huang Chao,Zhou Xuan,Han Runhua,Deng Yu,Zhou Shenghu 한국미생물·생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.10

Although rational genetic engineering is nowadays the favored method for microbial strain improvement, building up mutant libraries based on directed evolution for improvement is still in many cases the better option. In this regard, the demand for precise and efficient screening methods for mutants with high performance has stimulated the development of biosensor-based highthroughput screening strategies. Genetically encoded biosensors provide powerful tools to couple the desired phenotype to a detectable signal, such as fluorescence and growth rate. Herein, we review recent advances in engineering several classes of biosensors and their applications in directed evolution. Furthermore, we compare and discuss the screening advantages and limitations of two-component biosensors, transcription-factor-based biosensors, and RNA-based biosensors. Engineering these biosensors has focused mainly on modifying the expression level or structure of the biosensor components to optimize the dynamic range, specificity, and detection range. Finally, the applications of biosensors in the evolution of proteins, metabolic pathways, and genome-scale metabolic networks are described. This review provides potential guidance in the design of biosensors and their applications in improving the bioproduction of microbial cell factories through directed evolution.

Biosensors for rapid and sensitive detection of <i>Staphylococcus aureus</i> in food

Rubab, Momna,Shahbaz, Hafiz Muhammad,Olaimat, Amin N.,Oh, Deog-Hwan Elsevier 2018 Biosensors & bioelectronics Vol.105 No.-

<P><B>Abstract</B></P> <P>Foodborne illness outbreaks caused by the consumption of food contaminated with harmful bacteria has drastically increased in the past decades. Therefore, detection of harmful bacteria in the food has become an important factor for the recognition and prevention of problems associated with food safety and public health. <I>Staphylococcus aureus</I> is one of the most commonly isolated foodborne pathogen and it is considered as a major cause of foodborne illnesses worldwide. A number of different methods have been developed for the detection and identification of <I>S. aureus</I> in food samples. However, some of these methods are laborious and time-consuming and are not suitable for on-site applications. Therefore, it is highly important to develop rapid and more approachable detection methods. In the last decade, biosensors have gained popularity as an attractive alternative method and now considered as one of most rapid and on-site applicable methods. An overview of the biosensor based methods used for the detection of <I>S. aureus</I> is presented herein. This review focuses on the state-of-the-art biosensor methods towards the detection and quantification of <I>S. aureus,</I> and discusses the most commonly used biosensor methods based on the transducing mode, such as electrochemical, optical, and mass-based biosensors.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Recent advances in development of biosensors for detection of <I>S. aureus</I> are discussed. </LI> <LI> An overview of biosensors based on transducing mode is presented. </LI> <LI> Electrochemical, optical, and mass-based biosensors are mainly discussed. </LI> </UL> </P>

SWCNT GO/Co/chitosan 기반 바이오센서를 이용한 땀 속 포도당, 젖산 및 요소 검출

김동섭 ( Dong Sup Kim ),장윤원 ( Yun Won Jang ),정찬엽 ( Chan Yeop Jung ),신윤아 ( Yoona Shin ),이진영 ( Jinyoung Lee ) 한국키틴키토산학회 2024 한국키틴키토산학회지 Vol.29 No.1

본 연구에서 진행한 바이오센서는 인체의 땀에서의 대사물질을 검출하는 서로 다른 효소가 고정되어 있는 나노바이오센서로서, 그래핀 탄소 코팅된 단일벽 탄소나노튜브를 포함한다. 땀 속에 있는 타깃 대사 물질 검출용 센서 수용체로 포도당 산화효소 (Gox), 젖산 산화효소 (Lox), 유레이즈 (Urease)를 선택하였고, 이와 반응하는 기질인 Glucose, Lactic acid, Urea를 사용하였다. 실험 결과 대사물질의 산화효소 (Gox, Lox, Ure 등)를 고정화한 탄소나노튜브 기반 전극에 일정량의 대사물질 (포도당, 젖산, 우레아 등) 를 반응시킬 때 세 가지 대사물질 모두 저농도부터 고농도까지 측정됨을 확인할 수 있었다. 따라서 본 연구에서 개발한 바이오센서의 검출 효과가 있었음을 확인하였다. 본 실험은 세 개의 대사물에 대해서 독립적으로 실험을 진행했다. 순차적으로 인공 땀, 실제 인체 땀으로 실험하면 더 효과적인 바이오센서 개발이 될 수 있을 것이다. 추가적으로 이 연구를 바탕으로 탄소나노튜브 기반 전극을 활용하여 당 측정 바이오센서를 개발하고자 할 때 복수의 검출 대사물(포도당, 젖산 및 우레아)을 하나의 샘플에서 독립적으로 검출할 수 있는지에 대한 실험을 진행한다면 더 효과적인 바이오센서 개발을 할 수 있을 것이라고 생각한다. Recently, biosensor for detection of nutrients in human metabolic is one of attractive innovation four generation due to real time monitoring. It is necessary to develop a high-sensitive biosensor for detection of human organism components. In this study, Single walled carbon nanotube (SWCNT) based biosensor with high sensitivity was developed to detect glucose (Glu), lactic acid (Lac), and urea (Ure). Presented in this work is the development of a simple and highly sensitive D-glucose biosensor containing glucose oxidase (Gox), lactate oxidase (Lox), and Urease (Urs)-immobilized on SWNTs, respectively. Employing simple directed assembly and non-covalent functionlizationprocess these fabricated Gox, Lox, and Urs modified SWCNTs-based biosensors were designed with two electrode terminals to allow continuous resistance, cyclicvoltammetry and real time current response monitoring for the detection of Glu, Lac, and Ure, respectively. The developed biosensor had detection range in small size can make this biosensor potentially useful for in-vivo mode applications. SWCNT, 1-pyrenebutanoic acid, succinimide as electron transfer mediator, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxy succinimide (NHS) as linker, and three enzymes (Gox, Lox, and Urs) as acceptors, were step by step immobilized in the manufacturing process of biosensor. The developed biosensor has been confirmed to be capable of detecting three target metabolites (Glu, Lac, and Ure) in human components. Therefore, it is expected that this study can be used as a basic study for food assessment in large industry. Therefore, it is expected that this study can be used as a basic study for human body assessment in large industry.

CRISPR/dCas9-mediated biosensor for detection of tick-borne diseases

Koo, Bonhan,Kim, Da-eun,Kweon, Jiyeon,Jin, Choong Eun,Kim, Sung-Han,Kim, Yongsub,Shin, Yong Elsevier 2018 Sensors and actuators. B Chemical Vol.273 No.-

<P><B>Abstract</B></P> <P>Rapid and highly sensitive detection of biomolecules is greatly needed for pathogen diagnosis in clinical samples, but the method needs to be significantly improved in terms of sensitivity and specificity for actual use in clinical settings. Here, we report the development of an improved molecular diagnostics tool that utilizes CRISPR/dCas9-mediated biosensor that couples a nuclease inactivated Cas9 (dCas9) and single microring resonator biosensor, enables label-free and real-time detection of pathogenic DNA and RNA. We addressed the clinical utility of this CRISPR/dCas9-mediated biosensor in tick-borne illnesses including scrub typhus (ST) and severe fever with thrombocytopenia syndrome (SFTS), whose clinical presentations are too similar to be easily differentiated. By using CRISPR/dCas9-mediated biosensor, we achieved single molecule sensitivity for the detection of ST (0.54 aM) and SFTS (0.63 aM); this detection sensitivity is 100 times more sensitive than that of RT-PCR assay. Finally, CRISPR/dCas9-mediated biosensor was able to clearly distinguish between ST and SFTS in serum samples within 20 min. We believe that CRISPR/dCas9-mediated biosensor will be useful for rapid and accurate molecular diagnostic tool that is suitable for immediate clinical applications.</P> <P><B>Highlights</B></P> <P> <UL> <LI> An improved molecular diagnostics tool that utilizes CRISPR/dCas9-mediated biosensor. </LI> <LI> Couples a nuclease inactivated Cas9 (dCas9) and single microring resonator biosensor. </LI> <LI> Achieved single molecule sensitivity for the detection of ST (0.54 aM) and SFTS (0.63 aM). </LI> <LI> CRISPR/dCas9-mediated biosensor can detect pathogens in 20 min. </LI> </UL> </P>

Development of electrochemical biosensor for detection of pathogenic microorganism in Asian dust events

Yoo, Min-Sang,Shin, Minguk,Kim, Younghun,Jang, Min,Choi, Yoon-E,Park, Si Jae,Choi, Jonghoon,Lee, Jinyoung,Park, Chulhwan Elsevier 2017 CHEMOSPHERE - Vol.175 No.-

<P><B>Abstract</B></P> <P>We developed a single-walled carbon nanotubes (SWCNTs)-based electrochemical biosensor for the detection of <I>Bacillus subtilis,</I> one of the microorganisms observed in Asian dust events, which causes respiratory diseases such as asthma and pneumonia. SWCNTs plays the role of a transducer in biological antigen/antibody reaction for the electrical signal while 1-pyrenebutanoic acid succinimidyl ester (1-PBSE) and ant-<I>B. subtilis</I> were performed as a chemical linker and an acceptor, respectively, for the adhesion of target microorganism in the developed biosensor. The detection range (10<SUP>2</SUP>–10<SUP>10</SUP> CFU/mL) and the detection limit (10<SUP>2</SUP> CFU/mL) of the developed biosensor were identified while the response time was 10 min. The amount of target <I>B. subtilis</I> was the highest in the specificity test of the developed biosensor, compared with the other tested microorganisms (<I>Staphylococcus aureus</I>, <I>Flavobacterium psychrolimnae</I>, and <I>Aquabacterium commune</I>). In addition, target <I>B. subtilis</I> detected by the developed biosensor was observed by scanning electron microscope (SEM) analysis.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A SWCNTs-based biosensor was developed for the detection of <I>Bacillus subtilis.</I> </LI> <LI> The biosensor was composed of SWCNTs, 1-PBSE, and anti-<I>B. subtilis</I> antibody. </LI> <LI> The performance of the biosensor was assessed using a sensor sensitivity and specificity tests. </LI> <LI> The detection limit and detection range were 10<SUP>2</SUP> and 10<SUP>2</SUP>–10<SUP>10</SUP> CFU/mL, respectively. </LI> <LI> The detection time of the biosensor was identified as 10 min. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>