Protein adsorption/desorption and antibody binding stoichiometry on silicon interferometric biosensors examined with TOF-SIMS

Handle URI:
http://hdl.handle.net/10754/627335
Title:
Protein adsorption/desorption and antibody binding stoichiometry on silicon interferometric biosensors examined with TOF-SIMS
Authors:
Gajos, Katarzyna; Budkowski, Andrzej; Petrou, Panagiota; Pagkali, Varvara; Awsiuk, Kamil; Rysz, Jakub; Bernasik, Andrzej; Misiakos, Konstantinos; Raptis, Ioannis ( 0000-0001-5716-6157 ) ; Kakabakos, Sotirios
Abstract:
Time-of-flight secondary ion mass spectrometry has been employed to examine, with biomolecular discrimination, sensing arm areas (20 μm x 600 μm) of integrated onto silicon chips Mach-Zehnder interferometers aiming to optimize their biofunctionalization with regard to indirect immunochemical (competitive) detection of ochratoxin A. Sensing areas are examined after: modification with (3-aminopropyl)triethoxysilane, spotting of OTA-ovalbumin conjugate (probe) from solutions with different concentration, blocking with bovine serum albumin, reaction with OTA-specific mouse monoclonal antibody followed by goat anti-mouse IgG secondary antibody. Component mass loadings of all proteins involved in immunodetection are determined from TOF-SIMS micro-analysis combined with ellipsometry of planar surfaces. These data show that partial desorption of surface-bound probe and blocking protein takes place upon primary immunoreaction to a degree that depends on probe concentration in spotting solution. Taking into account this desorption, apparent binding stoichiometry of both antibodies in immune complexes formed onto chip surface is determined more accurately than the respective evaluation based on real-time sensor response. In addition, mass loadings for probe and secondary antibody is observed to saturate for optimum probe concentrations. Also, principal component analysis of TOF-SIMS data could resolve both immunoreactions and biofunctionalization and discriminate surfaces prepared with optimum probe concentrations from those prepared using suboptimum ones.
KAUST Department:
King Abdullah University of Science and Technology (KAUST), Core Labs, Thuwal, 23955-6900, Saudi Arabia.
Citation:
Gajos K, Budkowski A, Petrou P, Pagkali V, Awsiuk K, et al. (2018) Protein adsorption/desorption and antibody binding stoichiometry on silicon interferometric biosensors examined with TOF-SIMS. Applied Surface Science. Available: http://dx.doi.org/10.1016/j.apsusc.2018.03.029.
Publisher:
Elsevier BV
Journal:
Applied Surface Science
Issue Date:
5-Mar-2018
DOI:
10.1016/j.apsusc.2018.03.029
Type:
Article
ISSN:
0169-4332
Sponsors:
This work was supported by the EU funded project “FOODSNIFFER” (FP7-ICT8-318319). Data analysis was partially financed by Polish National Science Center (NCN) under Grant 2016/21/N/ST5/00880. The equipment used was purchased thanks to the financial support of the European Regional Development Fund (POIG.02.01.00-12-023/08).
Additional Links:
https://www.sciencedirect.com/science/article/pii/S0169433218306895
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Full metadata record

DC FieldValue Language
dc.contributor.authorGajos, Katarzynaen
dc.contributor.authorBudkowski, Andrzejen
dc.contributor.authorPetrou, Panagiotaen
dc.contributor.authorPagkali, Varvaraen
dc.contributor.authorAwsiuk, Kamilen
dc.contributor.authorRysz, Jakuben
dc.contributor.authorBernasik, Andrzejen
dc.contributor.authorMisiakos, Konstantinosen
dc.contributor.authorRaptis, Ioannisen
dc.contributor.authorKakabakos, Sotiriosen
dc.date.accessioned2018-03-15T11:35:53Z-
dc.date.available2018-03-15T11:35:53Z-
dc.date.issued2018-03-05en
dc.identifier.citationGajos K, Budkowski A, Petrou P, Pagkali V, Awsiuk K, et al. (2018) Protein adsorption/desorption and antibody binding stoichiometry on silicon interferometric biosensors examined with TOF-SIMS. Applied Surface Science. Available: http://dx.doi.org/10.1016/j.apsusc.2018.03.029.en
dc.identifier.issn0169-4332en
dc.identifier.doi10.1016/j.apsusc.2018.03.029en
dc.identifier.urihttp://hdl.handle.net/10754/627335-
dc.description.abstractTime-of-flight secondary ion mass spectrometry has been employed to examine, with biomolecular discrimination, sensing arm areas (20 μm x 600 μm) of integrated onto silicon chips Mach-Zehnder interferometers aiming to optimize their biofunctionalization with regard to indirect immunochemical (competitive) detection of ochratoxin A. Sensing areas are examined after: modification with (3-aminopropyl)triethoxysilane, spotting of OTA-ovalbumin conjugate (probe) from solutions with different concentration, blocking with bovine serum albumin, reaction with OTA-specific mouse monoclonal antibody followed by goat anti-mouse IgG secondary antibody. Component mass loadings of all proteins involved in immunodetection are determined from TOF-SIMS micro-analysis combined with ellipsometry of planar surfaces. These data show that partial desorption of surface-bound probe and blocking protein takes place upon primary immunoreaction to a degree that depends on probe concentration in spotting solution. Taking into account this desorption, apparent binding stoichiometry of both antibodies in immune complexes formed onto chip surface is determined more accurately than the respective evaluation based on real-time sensor response. In addition, mass loadings for probe and secondary antibody is observed to saturate for optimum probe concentrations. Also, principal component analysis of TOF-SIMS data could resolve both immunoreactions and biofunctionalization and discriminate surfaces prepared with optimum probe concentrations from those prepared using suboptimum ones.en
dc.description.sponsorshipThis work was supported by the EU funded project “FOODSNIFFER” (FP7-ICT8-318319). Data analysis was partially financed by Polish National Science Center (NCN) under Grant 2016/21/N/ST5/00880. The equipment used was purchased thanks to the financial support of the European Regional Development Fund (POIG.02.01.00-12-023/08).en
dc.publisherElsevier BVen
dc.relation.urlhttps://www.sciencedirect.com/science/article/pii/S0169433218306895en
dc.rightsNOTICE: this is the author’s version of a work that was accepted for publication in Applied Surface Science. 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 Applied Surface Science, [, , (2018-03-05)] DOI: 10.1016/j.apsusc.2018.03.029 . © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjecton-chip biosensoren
dc.subjectTime-of-Flight Secondary Ion Mass Spectrometryen
dc.subjectMulti-component layersen
dc.subjectBiofunctionalizationen
dc.subjectIndirect immunoassayen
dc.subjectBinding stoichiometryen
dc.titleProtein adsorption/desorption and antibody binding stoichiometry on silicon interferometric biosensors examined with TOF-SIMSen
dc.typeArticleen
dc.contributor.departmentKing Abdullah University of Science and Technology (KAUST), Core Labs, Thuwal, 23955-6900, Saudi Arabia.en
dc.identifier.journalApplied Surface Scienceen
dc.eprint.versionPost-printen
dc.contributor.institutionM. Smoluchowski Institute of Physics, Jagiellonian University, Łojasiewicza 11, 30-348 Kraków, Polanden
dc.contributor.institutionInstitute of Nuclear & Radiological Sciences & Technology, Energy & Safety NCSR Demokritos, P. Grigoriou & Neapoleos St, Aghia Paraksevi 15310, Athens, Greeceen
dc.contributor.institutionAcademic Centre for Materials and Nanotechnology, AGH University of Science and Technology, Mickiewicza 30, 30-059 Kraków, Polanden
dc.contributor.institutionFaculty of Physics and Applied Computer Science, AGH University of Science and Technology, Mickiewicza 30, 30-059 Kraków, Polanden
dc.contributor.institutionInstitute of Nanoscience and Nanotechnology, NCSR Demokritos, P. Grigoriou & Neapoleos St, Aghia Paraskevi 15310, Athens, Greeceen
kaust.authorRaptis, Ioannisen
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