Optimization and characterization of covalent immobilization of glucose oxidase for bioelectronic devices

Wang, Xue; Kim, Sung‐Bae; Khang, Dongwoo; Kim, Hyug-Han; Kim, Chang-Joon

doi:10.1016/j.bej.2016.03.016

Cited by 19 publications

(5 citation statements)

References 52 publications

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“…Glucose oxidase from Aspergillus niger (GOx, type X-S, 100,000-250,000 units/g, Sigma-Aldrich G7141) was covalently bonded to the PNU-VACNT array using 1-ethyl-3-(-3dimethylaminopropyl) carbodiimide (EDC, Sigma-Aldrich E7750) and N-hydroxysuccinimide (NHS, Sigma-Aldrich 130672) chemistry following approximately the conditions that were optimized by Wang, et al (see Figure 2C). 38 The PNU-VACNT array was incubated in 5 mL of 50 mM EDC and 400 mM NHS in 0.1 M 2-(N-morpholino) ethanesulfonic acid (MES, pH 4.7, Thermo Scientific 28390) for 90 minutes to allow the carboxyl groups on the VACNTs to react with EDC and to form NHS esters in preparation for GOx coupling. The sample was then rinsed with ultrapure water and then placed in a 5 mL phosphate buffered saline (PBS, pH 7.4) solution with 50 mg of GOx (10 mg/mL).…”

Section: Covalent (Edc/nhs) Bindingmentioning

confidence: 99%

Electrochemical Glucose Sensors Enhanced by Methyl Viologen and Vertically Aligned Carbon Nanotube Channels

Brownlee

Bahari

Harb

et al. 2018

ACS Appl. Mater. Interfaces

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Freestanding, vertically aligned carbon nanotubes (VACNTs) were patterned into 16 μm diameter microchannel arrays for flow-through electrochemical glucose sensing. Non-enzymatic sensing of glucose was achieved by the chemical reaction of glucose with methyl viologen (MV) at an elevated temperature and pH (0.1 M NaOH), followed by the electrochemical reaction of reduced-MV with the VACNT surface. The MV sensor required no functionalization (including no metal) and was able to produce on average 3.4 electrons per glucose molecule. The current density of the MV sensor was linear with both flow rate and glucose concentration. Challenges with interference chemicals were mitigated by operating at a low potential of -0.2 V vs Ag/AgCl. As a comparison, enzymatic VACNT sensors with platinum nano-urchins were functionalized with glucose oxidase by covalent binding (1-ethyl-3-(-3-dimethylaminopropyl)carbodiimide/ N-hydroxysuccinimide) or by polymer entrapment [poly(3,4-ethylene-dioxythiophene)] and operated in phosphate buffered saline. With normalization by the overall cross-sectional area of the flow (0.713 cm), the sensitivity of the MV, enzyme-in-solution, and covalent sensors were 45.93, 18.77, and 1.815 mA cm mM, respectively. Corresponding limits of detection were 100, 194, and 311 nM glucose. The linear sensing ranges for the sensors were 250 nM to 200 μM glucose for the MV sensor, 500 nM to 200 μM glucose for the enzyme-in-solution sensor, and 1 μM to 6 mM glucose for the covalent sensor. The flow cell and sensor cross-sectional area were scaled down (0.020 cm) to enable detection from 200 μL of glucose with MV by flow injection analysis. The sensitivity of the small MV sensor was 5.002 mA cm mM, with a limit of detection of 360 nM glucose and a linear range up to at least 150 μM glucose. The small MV sensor has the potential to measure glucose levels found in 200 μL of saliva.

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Section: Covalent (Edc/nhs) Bindingmentioning

confidence: 99%

Electrochemical Glucose Sensors Enhanced by Methyl Viologen and Vertically Aligned Carbon Nanotube Channels

Brownlee

Bahari

Harb

et al. 2018

ACS Appl. Mater. Interfaces

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“…A list of major nano-materials, CNTs, and carbon nano-fibers (CNFs) used as electrical connectors between the electrode and the redox center, are given in Table 3 . Wang X. et al ( 2016 ) recently reported optimization and characterization of covalent immobilization of GOD using multi-walled carbon nano-tubes (MWCNTs) to maximize the loading of GOD, thereby, increasing the longevity of electric power or sensing signals.…”

Section: Applications Of God In Various Sectorsmentioning

confidence: 99%

Improvement Strategies, Cost Effective Production, and Potential Applications of Fungal Glucose Oxidase (GOD): Current Updates

et al. 2017

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Fungal glucose oxidase (GOD) is widely employed in the different sectors of food industries for use in baking products, dry egg powder, beverages, and gluconic acid production. GOD also has several other novel applications in chemical, pharmaceutical, textile, and other biotechnological industries. The electrochemical suitability of GOD catalyzed reactions has enabled its successful use in bioelectronic devices, particularly biofuel cells, and biosensors. Other crucial aspects of GOD such as improved feeding efficiency in response to GOD supplemental diet, roles in antimicrobial activities, and enhancing pathogen defense response, thereby providing induced resistance in plants have also been reported. Moreover, the medical science, another emerging branch where GOD was recently reported to induce several apoptosis characteristics as well as cellular senescence by downregulating Klotho gene expression. These widespread applications of GOD have led to increased demand for more extensive research to improve its production, characterization, and enhanced stability to enable long term usages. Currently, GOD is mainly produced and purified from Aspergillus niger and Penicillium species, but the yield is relatively low and the purification process is troublesome. It is practical to build an excellent GOD-producing strain. Therefore, the present review describes innovative methods of enhancing fungal GOD production by using genetic and non-genetic approaches in-depth along with purification techniques. The review also highlights current research progress in the cost effective production of GOD, including key advances, potential applications and limitations. Therefore, there is an extensive need to commercialize these processes by developing and optimizing novel strategies for cost effective GOD production.

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“…Silicon photonic resonator-based devices rely on the resonant property of light signal transmission. The resonant wavelength shift of light through the resonator is responsive to the effective refractive index change that occurs through biomaterial binding on the waveguide surface, such as antibody-antigen [11][12][13] or enzyme-substrate [14,15]. Therefore, the coating of a bioreceptor-such as an antibody or enzyme-on silicon is the essential stage where a high density of protein molecules must be immobilized onto silicon surface uniformly.…”

Section: Introductionmentioning

confidence: 99%

“…Hydroxyl groups (-OH) are used by the silanization step by the hydrolysis of ethoxy (-C 2 H 5 ) groups from APTES, leading to the formation of silanols (Si-OH). APTES silanols can condense with surface silanols and express amine groups, which can further crosslink with a bioreceptor protein (e.g., antibody) using a GA linker [11][12][13][14][15][16][17][18][19]. There is a potential issue with crosslinking by GA since both functional groups (aldehydes) of GA molecules are identical, and nonspecific to proteins.…”

Section: Introductionmentioning

confidence: 99%

Novel Rapid Protein Coating Technique for Silicon Photonic Biosensor to Improve Surface Morphology and Increase Bioreceptor Density

et al. 2021

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Silicon photonic devices with either silicon or silicon nitride waveguides have increasingly been used in many applications besides communications, especially as sensors in label-free biosensing, where guided light signals are affected by biorecognition molecules immobilized on the surface. The coating of protein (i.e., bioreceptors) by biochemical process on the waveguide surface is a crucial step in creating a functionalized device that can be used for biosensing. As a conventional method that uses 3-aminopropryltriethoxysilane (APTES) and glutaraldehyde (GA), the APTES-GA method has the limitation of using a GA crosslink, of which the two functional groups can bind to nonspecific proteins, causing irregular binding. In this study, we proposed a new coating technique to avoid such problem by applying APTES silanization with 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide (EDC)-N-hydroxysuccinimide (NHS) protein crosslink, denoted by the APTES-(EDC/NHS) method. The EDC/NHS reaction was shown to be able to immobilize protein in ordered orientation due to consistent arrangement between a carboxylic group of protein molecules and an amine group of covalent-linked APTES on surface. By applying APTES silanization, we circumvented the use of hazardous cleaning agent in the conventional EDC/NHS technique. Several surface characterization techniques were carried out to assess and compare the two biocoating techniques, including scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), spectroscopic ellipsometry (SE), and atomic force microscopy (AFM). On silicon, the results of antihuman TNF-alpha antibody coating showed that the proposed APTES-(EDC/NHS) technique has better repeatability in terms of less roughness of the coated protein at 1.5 nm compared with 6.3 nm, due to the ordered arrangement of coated antibody molecules. On a silicon nitride waveguide device, the proposed APTES-(EDC/NHS) technique exhibits dense antibody immobilization on a waveguide in SEM images due to stable amide bond formation via EDC/NHS crosslink mechanism. The specificity of the immobilized antibodies was confirmed by enzyme-linked immunosorbent assays (ELISA), with an average optical density at 450 nm of 0.175 ± 0.01 compared with 0.064 ± 0.009 of negative control. The proposed technique also reduced the overall process time since proteins are crosslinked to the silanized waveguide surface in a single step.

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Optimization and characterization of covalent immobilization of glucose oxidase for bioelectronic devices

Cited by 19 publications

References 52 publications

Electrochemical Glucose Sensors Enhanced by Methyl Viologen and Vertically Aligned Carbon Nanotube Channels

Electrochemical Glucose Sensors Enhanced by Methyl Viologen and Vertically Aligned Carbon Nanotube Channels

Improvement Strategies, Cost Effective Production, and Potential Applications of Fungal Glucose Oxidase (GOD): Current Updates

Novel Rapid Protein Coating Technique for Silicon Photonic Biosensor to Improve Surface Morphology and Increase Bioreceptor Density

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