Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensor
The commercial success of paper-based microfluidic biosensor devices with enzyme as recognition elements may be achieved greatly by improving the enzyme stability, averting the coffee-ring effect on the detection zone, and making it a stand-alone system. We have addressed these issues here by introd...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | Biosensors and Bioelectronics: X |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590137022000425 |
| _version_ | 1811281172981874688 |
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| author | Phurpa Dema Thungon Pooja Rani Kuri Vinay Bachu Pranab Goswami |
| author_facet | Phurpa Dema Thungon Pooja Rani Kuri Vinay Bachu Pranab Goswami |
| author_sort | Phurpa Dema Thungon |
| collection | DOAJ |
| description | The commercial success of paper-based microfluidic biosensor devices with enzyme as recognition elements may be achieved greatly by improving the enzyme stability, averting the coffee-ring effect on the detection zone, and making it a stand-alone system. We have addressed these issues here by introducing silk-fibroin (SF) film for immobilizing enzymes and chromogenic reagent (ABTS), and wave-design microfluidic channels in the chromatographic paper for developing a hybrid microfluidic paper-based analytical device (μPAD), for methanol detection through peroxidase reaction. The activity of alcohol oxidase (AOx), used as a biorecognition element, could be wholly retained in a non-dissolvable SF film (pore size <3.5 μm) until 40 days of storage at room temperature (RT) by reducing oxygen permeability to the film. Similarly, ABTS in a dissolvable SF film was protected from air-oxidation even up to two months of storage at RT. The detection approach exploited purple color as a high contrast response signal generated from ABTS di-cation formed from the reaction of SF protein with the ABTS radicals generated from the substrate-dependent peroxidase reaction. The wave-designed microfluidic channels could significantly reduce the coffee ring effect in the detection film. The bi-enzyme SF films prepared by co-immobilizing AOx with peroxidase (HRP) offered high sensitivity to the device with a detection limit (LoD) of 1 ± 0.05 mM and a dynamic range of 1 mM–2 M for methanol. The stand-alone hybrid μPAD biosensor for methanol has a high potential for commercial applications. |
| first_indexed | 2024-04-13T01:27:53Z |
| format | Article |
| id | doaj.art-9af5a54df67b434cb8d763cdc12ff30d |
| institution | Directory Open Access Journal |
| issn | 2590-1370 |
| language | English |
| last_indexed | 2024-04-13T01:27:53Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biosensors and Bioelectronics: X |
| spelling | doaj.art-9af5a54df67b434cb8d763cdc12ff30d2022-12-22T03:08:35ZengElsevierBiosensors and Bioelectronics: X2590-13702022-09-0111100147Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensorPhurpa Dema Thungon0Pooja Rani Kuri1Vinay Bachu2Pranab Goswami3Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Assam, 781039, IndiaDepartment of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Assam, 781039, IndiaDepartment of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Assam, 781039, IndiaCorresponding author.; Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Assam, 781039, IndiaThe commercial success of paper-based microfluidic biosensor devices with enzyme as recognition elements may be achieved greatly by improving the enzyme stability, averting the coffee-ring effect on the detection zone, and making it a stand-alone system. We have addressed these issues here by introducing silk-fibroin (SF) film for immobilizing enzymes and chromogenic reagent (ABTS), and wave-design microfluidic channels in the chromatographic paper for developing a hybrid microfluidic paper-based analytical device (μPAD), for methanol detection through peroxidase reaction. The activity of alcohol oxidase (AOx), used as a biorecognition element, could be wholly retained in a non-dissolvable SF film (pore size <3.5 μm) until 40 days of storage at room temperature (RT) by reducing oxygen permeability to the film. Similarly, ABTS in a dissolvable SF film was protected from air-oxidation even up to two months of storage at RT. The detection approach exploited purple color as a high contrast response signal generated from ABTS di-cation formed from the reaction of SF protein with the ABTS radicals generated from the substrate-dependent peroxidase reaction. The wave-designed microfluidic channels could significantly reduce the coffee ring effect in the detection film. The bi-enzyme SF films prepared by co-immobilizing AOx with peroxidase (HRP) offered high sensitivity to the device with a detection limit (LoD) of 1 ± 0.05 mM and a dynamic range of 1 mM–2 M for methanol. The stand-alone hybrid μPAD biosensor for methanol has a high potential for commercial applications.http://www.sciencedirect.com/science/article/pii/S2590137022000425Silk fibroinAlcohol oxidaseABTSPeroxidaseEnzyme stabilityMethanol biosensors |
| spellingShingle | Phurpa Dema Thungon Pooja Rani Kuri Vinay Bachu Pranab Goswami Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensor Biosensors and Bioelectronics: X Silk fibroin Alcohol oxidase ABTS Peroxidase Enzyme stability Methanol biosensors |
| title | Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensor |
| title_full | Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensor |
| title_fullStr | Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensor |
| title_full_unstemmed | Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensor |
| title_short | Silk-fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase-based μPAD methanol biosensor |
| title_sort | silk fibroin film as enzyme stabilizing material and optical signal transducer for developing alcohol oxidase based μpad methanol biosensor |
| topic | Silk fibroin Alcohol oxidase ABTS Peroxidase Enzyme stability Methanol biosensors |
| url | http://www.sciencedirect.com/science/article/pii/S2590137022000425 |
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