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dc.contributor.authorKulsharova, Gulsim
dc.contributor.authorDimov, Nikolay
dc.contributor.authorMarques, Marco P.C.
dc.contributor.authorSzita, Nicolas
dc.contributor.authorBaganz, Frank
dc.date.accessioned2018-09-25T09:13:25Z
dc.date.available2018-09-25T09:13:25Z
dc.date.issued2017-12-11
dc.identifier.citationKulsharova , G , Dimov , N , Marques , M P C , Szita , N & Baganz , F 2017 , ' Simplified immobilisation method for histidine-tagged enzymes in poly(methyl methacrylate) microfluidic devices ' , New Biotechnology . https://doi.org/10.1016/j.nbt.2017.12.004
dc.identifier.issn1871-6784
dc.identifier.otherPURE: 13219573
dc.identifier.otherPURE UUID: 3ffdff33-5794-4cad-b693-948ac2d5f4a0
dc.identifier.otherScopus: 85038021922
dc.identifier.otherORCID: /0000-0002-2873-1505/work/63687438
dc.identifier.urihttp://hdl.handle.net/2299/20644
dc.descriptionArticle in press. Kulsharova, G., New BIOTECHNOLOGY (2017), https://doi.org/10.1016/j.nbt.2017.12.004
dc.description.abstractPoly(methyl methacrylate) (PMMA) microfluidic devices have become promising platforms for a wide range of applications. Here we report a simple method for immobilising histidine-tagged enzymes suitable for PMMA microfluidic devices. The 1-step-immobilisation described is based on the affinity of the His-tag/Ni-NTA interaction and does not require prior amination of the PMMA surface, unlike many existing protocols. We compared it with a 3-step immobilisation protocol involving amination of PMMA and linking NTA via a glutaraldehyde cross-linker. These methods were applied to immobilise transketolase (TK) in PMMA microfluidic devices. Binding efficiency studies showed that about 15% of the supplied TK was bound using the 1-step method and about 26% of the enzyme was bound by the 3-step method. However, the TK-catalysed reaction producing l-erythrulose performed in microfluidic devices showed that specific activity of TK in the device utilising the 1-step immobilisation method was approximately 30% higher than that of its counterpart. Reusability of the microfluidic device produced via the 1-step method was tested for three cycles of enzymatic reaction and at least 85% of the initial productivity was maintained. The device could be operated for up to 40 h in a continuous flow and on average 70% of the initial productivity was maintained. The simplified immobilisation method required fewer chemicals and less time for preparation of the immobilised microfluidic device compared to the 3-step method while achieving higher specific enzyme activity. The method represents a promising approach for the development of immobilised enzymatic microfluidic devices and could potentially be applied to combine protein purification with immobilisation.en
dc.language.isoeng
dc.relation.ispartofNew Biotechnology
dc.subjectEnzyme immobilisation
dc.subjectHistidine-tagged enzyme
dc.subjectMicrofluidics
dc.subjectPoly(methyl methacrylate) (PMMA)
dc.subjectTransketolase
dc.subjectBiotechnology
dc.subjectBioengineering
dc.subjectMolecular Biology
dc.titleSimplified immobilisation method for histidine-tagged enzymes in poly(methyl methacrylate) microfluidic devicesen
dc.contributor.institutionSchool of Physics, Engineering & Computer Science
dc.description.statusPeer reviewed
dc.identifier.urlhttp://www.scopus.com/inward/record.url?scp=85038021922&partnerID=8YFLogxK
rioxxterms.versionAM
rioxxterms.versionVoR
rioxxterms.versionofrecordhttps://doi.org/10.1016/j.nbt.2017.12.004
rioxxterms.typeJournal Article/Review
herts.preservation.rarelyaccessedtrue


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