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Low back-pressure hierarchically structured multichannel microfluidic bioreactors for rapid protein digestion - proof of concept

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dc.contributor.author Szymanska, Katarzyna
dc.contributor.author Peitrowska, Monika
dc.contributor.author Kocurek, Jacek
dc.contributor.author Maresz, Katarzyna
dc.contributor.author Koreniuk, Agnieszka
dc.contributor.author Mrowiec-Białoń, Julita
dc.contributor.author Widłak, Piotr
dc.contributor.author Magner, Edmond
dc.contributor.author Jarzębski, Andrzej
dc.date.accessioned 2016-06-20T14:08:32Z
dc.date.issued 2016
dc.identifier.uri http://hdl.handle.net/10344/5077
dc.description peer-reviewed en_US
dc.description.abstract A novel, easy-to-fabricate monolithic enzymatic microreactor with a hierarchical, torturous structure of flow-through channels of micrometric sizes and large mesopores was shown to enable rapid and very efficient digestion of proteins at high yields and exceptionally low back-pressures. Four silica monoliths with bi-modal 3D pore structure in micrometer and nanometer size scales were synthesized and characterized for structural and flow properties. The monolith with the highest total pore volume (4 cm(3)/g) and flow through channels 20-30 mu m in size, was further functionalized with trypsin to obtain multichannel immobilized enzyme (proteolytic) reactor (IMER). The value of permeability coefficient K evaluated for water (similar to 2.0 . 10(-11)) was found to be two orders of magnitude higher in the novel reactor than reported before for high-performance IMERs, enabling the flow rates of 750 mL/cm(2) min at pressure gradients of 64 kPa/cm. Very high practical potentials of the novel microbioreactor were demonstrated in the proteolysis of cytochrome c (Cyt-c) and myoglobin (Myo), without any earlier pretreatment. MALDI-TOF/TOF mass spectrometry analysis of sequence coverage was high: 70% (Cyt-c) and 90% (Myo) for 24 min digestion, and 39% (Cyt-c) and 53% (Myo) when the proteolysis time was reduced to 2.4 min. The proposed microreactors make full use of all advantages of microfuidic devices and mesoporous biocatalysts, and offer exceptional possibilities for biochemical/proteolytic applications in both large (production) and small (analytical) scales. (C) 2015 Elsevier B.V. All rights reserved. en_US
dc.language.iso eng en_US
dc.publisher Elsevier en_US
dc.relation.ispartofseries Chemical Engineering Journal;287, pp. 148-154
dc.relation.uri http://dx.doi.org/10.1016/j.cej.2015.10.120
dc.rights This is the author’s version of a work that was accepted for publication in Chemical Engineering Journal. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Chemical Engineering Journal, 287, pp. 148-154, http://dx.doi.org/10.1016/j.cej.2015.10.120 en_US
dc.subject continuous protein digestion en_US
dc.subject immobilized enzyme reactor en_US
dc.subject microfluidic reactor en_US
dc.title Low back-pressure hierarchically structured multichannel microfluidic bioreactors for rapid protein digestion - proof of concept en_US
dc.type info:eu-repo/semantics/article en_US
dc.type.supercollection all_ul_research en_US
dc.type.supercollection ul_published_reviewed en_US
dc.date.updated 2016-06-20T13:48:35Z
dc.description.version ACCEPTED
dc.identifier.doi 10.1016/j.cej.2015.10.120
dc.contributor.sponsor National Science Center (NSC) of Poland en_US
dc.contributor.sponsor National Science Center (NCN) of Poland en_US
dc.relation.projectid DEC-2013/09/D/ST8/04002
dc.date.embargoEndDate 2017-11-14
dc.embargo.terms 2017-11-14 en_US
dc.rights.accessrights info:eu-repo/semantics/openAccess en_US
dc.internal.rssid 1630260
dc.internal.copyrightchecked Yes
dc.description.status peer-reviewed


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