Immobilization of proteolytic enzymes on replica-molded thiol-ene micropillar reactors via thiol-gold interaction

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dc.contributor Aalto-yliopisto fi
dc.contributor Aalto University en
dc.contributor.author Tähkä, Sari
dc.contributor.author Sarfraz, Jawad
dc.contributor.author Urvas, Lauri
dc.contributor.author Provenzani, Riccardo
dc.contributor.author Wiedmer, Susanne K.
dc.contributor.author Peltonen, Jouko
dc.contributor.author Jokinen, Ville
dc.contributor.author Sikanen, Tiina
dc.date.accessioned 2019-05-06T09:09:22Z
dc.date.available 2019-05-06T09:09:22Z
dc.date.issued 2019-03-21
dc.identifier.citation Tähkä , S , Sarfraz , J , Urvas , L , Provenzani , R , Wiedmer , S K , Peltonen , J , Jokinen , V & Sikanen , T 2019 , ' Immobilization of proteolytic enzymes on replica-molded thiol-ene micropillar reactors via thiol-gold interaction ' ANALYTICAL AND BIOANALYTICAL CHEMISTRY . https://doi.org/10.1007/s00216-019-01674-9 en
dc.identifier.issn 1618-2642
dc.identifier.issn 1618-2650
dc.identifier.other PURE UUID: 3c5cf1cf-aef8-4f73-a243-c5f94217f3bc
dc.identifier.other PURE ITEMURL: https://research.aalto.fi/en/publications/immobilization-of-proteolytic-enzymes-on-replicamolded-thiolene-micropillar-reactors-via-thiolgold-interaction(3c5cf1cf-aef8-4f73-a243-c5f94217f3bc).html
dc.identifier.other PURE LINK: http://www.scopus.com/inward/record.url?scp=85063231210&partnerID=8YFLogxK
dc.identifier.other PURE FILEURL: https://research.aalto.fi/files/32889239/CHEM_T_hk_et_al_Immobilization_of_proteolytic_enzymes_2019_Analytical_and_Bioanalytical_Chemistry.pdf
dc.identifier.uri https://aaltodoc.aalto.fi/handle/123456789/37641
dc.description | openaire: EC/H2020/| 311705/EU//CUMTAS
dc.description.abstract We introduce rapid replica molding of ordered, high-aspect-ratio, thiol-ene micropillar arrays for implementation of microfluidic immobilized enzyme reactors (IMERs). By exploiting the abundance of free surface thiols of off-stoichiometric thiol-ene compositions, we were able to functionalize the native thiol-ene micropillars with gold nanoparticles (GNPs) and these with proteolytic α-chymotrypsin (CHT) via thiol-gold interaction. The micropillar arrays were replicated via PDMS soft lithography, which facilitated thiol-ene curing without the photoinitiators, and thus straightforward bonding and good control over the surface chemistry (number of free surface thiols). The specificity of thiol-gold interaction was demonstrated over allyl-rich thiol-ene surfaces and the robustness of the CHT-IMERs at different flow rates and reaction temperatures using bradykinin hydrolysis as the model reaction. The product conversion rate was shown to increase as a function of decreasing flow rate (increasing residence time) and upon heating of the IMER to physiological temperature. Owing to the effective enzyme immobilization onto the micropillar array by GNPs, no further purification of the reaction solution was required prior to mass spectrometric detection of the bradykinin hydrolysis products and no clogging problems, commonly associated with conventional capillary packings, were observed. The activity of the IMER remained stable for at least 1.5 h (continuous use), suggesting that the developed protocol may provide a robust, new approach to implementation of IMER technology for proteomics research. [Figure not available: see fulltext.]. en
dc.format.mimetype application/pdf
dc.language.iso en en
dc.publisher SPRINGER HEIDELBERG
dc.relation info:eu-repo/grantAgreement/EC/H2020/| 311705/EU//CUMTAS
dc.relation.ispartofseries ANALYTICAL AND BIOANALYTICAL CHEMISTRY en
dc.rights openAccess en
dc.subject.other Analytical Chemistry en
dc.subject.other Biochemistry en
dc.subject.other 215 Chemical engineering en
dc.title Immobilization of proteolytic enzymes on replica-molded thiol-ene micropillar reactors via thiol-gold interaction en
dc.type A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä fi
dc.description.version Peer reviewed en
dc.contributor.department University of Helsinki
dc.contributor.department Åbo Akademi University
dc.contributor.department Department of Chemistry and Materials Science
dc.subject.keyword Enzyme immobilization
dc.subject.keyword Gold nanoparticles
dc.subject.keyword Mass spectrometry
dc.subject.keyword Microfluidics
dc.subject.keyword Microreactors
dc.subject.keyword Thiol-enes
dc.subject.keyword Analytical Chemistry
dc.subject.keyword Biochemistry
dc.subject.keyword 215 Chemical engineering
dc.identifier.urn URN:NBN:fi:aalto-201905062761
dc.identifier.doi 10.1007/s00216-019-01674-9
dc.type.version publishedVersion


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