Indirect immunoassay on functionalized silicon surface: Molecular arrangement, composition and orientation examined step-by-step with multi-technique and multivariate analysis

Gajos, Katarzyna; Budkowski, Andrzej; Pagkali, Varvara; Petrou, Panagiota; Biernat, Monika; Awsiuk, Kamil; Rysz, Jakub; Bernasik, Andrzej; Μισιακός, Κωνσταντίνος; Raptis, Ioannis; Kakabakos, Sotirios

doi:10.1016/j.colsurfb.2016.11.009

Cited by 14 publications

(28 citation statements)

References 32 publications

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“…It is this parameter that is optimized in this work. A similar analysis has been previously performed on planar substrates imitating the MZI transducer [20], and the new findings are discussed in relation to those. To optimize the biofunctionalization procedure for efficient detection of analyte, the on-chip sensors where the analyte-protein conjugate (probe) has been immobilized from solutions with different concentration were examined following a strategy introduced recently [25].…”

Section: Introductionsupporting

confidence: 62%

“…Additionally, planar Si 3 N 4 surfaces were prepared and functionalized as the silicon chips with the integrated MZIs to be used for ellipsometry measurements [20].…”

Section: Biosensor Surface Functionalization and Assay Protocolmentioning

confidence: 99%

“…(Sentech Instruments GmBH) Spectroscopic Ellipsometer working over a wavelength range of 320-700 nm and at a fixed angle of incidence equal to 70º was used. Results were analyzed with the SpectraRay 3 software assuming the Cauchy dispersion formula and three-layer model consisted of silicon substrate/Si 3 N 4 /mixed SiO 2 and APTES/protein overlay as described before [20].…”

Section: Ellipsometry Measurementsmentioning

confidence: 99%

“…Also, partial protein desorption during primary immunoreaction, concluded in [20], is shown here to depend on the probe spotting concentration, confirming the generality of this recently reported phenomenon [25]. Third, the molecular orientation of immunoadsorbed primary (side-on) and secondary antibody (headon) [20] corresponds well to their binding stoichiometry in the immune complexes formed onto the MZI transducer (with the ratio of ~1.3 for primary antibody to probe, and ~2.5 for secondary to primary antibody), evaluated here with TOF-SIMS more accurately than estimated from sensor response. is observed for the OTA-OVA probe (a).…”

Section: The Examination Of Microstructured Biosensors Extended From mentioning

confidence: 99%

“…Recently, Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) has been recognized as a powerful technique to analyze biomolecular overlayers on solid supports [17], including biosensor surfaces, due to chemical specificity and surface sensitivity it exhibits. Indeed, TOF-SIMS has been successfully employed to discriminate between different molecules immobilized on the different surfaces and in particular to detect different proteins based in differences in their amino acids composition [14,[18][19][20][21][22][23]. Among the reported applications of TOF-SIMS technique for biosensor surface analysis the examination of microstructured interferometric biosensors based on Mach-Zehnder interferometers monolithically integrated on silicon chips modified following a multi-step protocol has been demonstrated [19].…”

Section: Introductionmentioning

confidence: 99%

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

Gajos

Budkowski

Petrou

et al. 2018

Applied Surface Science

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Highlights  Examining microstructured sensing areas of interferometric biosensors on silicon chip  Step-by-step surface modifications determined with biomolecular discrimination  Surface density of probe, blocking protein, primary and secondary antibody evaluated  Desorption upon primary immunoreaction depends on spotting concentration of probe  Binding stoichiometry evaluated more accurately than estimated from sensor response

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Section: Introductionsupporting

confidence: 62%

“…Additionally, planar Si 3 N 4 surfaces were prepared and functionalized as the silicon chips with the integrated MZIs to be used for ellipsometry measurements [20].…”

Section: Biosensor Surface Functionalization and Assay Protocolmentioning

confidence: 99%

Section: Ellipsometry Measurementsmentioning

confidence: 99%

Section: The Examination Of Microstructured Biosensors Extended From mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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

Gajos

Budkowski

Petrou

et al. 2018

Applied Surface Science

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Using a lactadherin-immobilized silicon surface for capturing and monitoring plasma microvesicles as a foundation for diagnostic device development

et al. 2020

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Microvesicles (MVs) are found in several types of body fluids and are promising disease biomarkers and therapeutic targets. This study aimed to develop a novel biofunctionalized surface for binding plasma microvesicles (PMVs) based on a lab-on-a-chip (LOC) approach. A new lactadherin (LACT)-functionalized surface was prepared and examined for monitoring PMVs. Moreover, two different strategies of LACT immobilization on a silicon surface were applied to compare different LACT orientations. A higher PMV to LACT binding efficiency was observed for LACT bonded to an αvβ3 integrin–functionalized surface compared with that for LACT directly bonded to a glutaraldehyde-modified surface. Effective binding of PMVs and its components for both LACT immobilization strategies was confirmed using spectral ellipsometry and time-of-flight secondary ion mass spectrometry methods. The proposed PMV capturing system can be used as a foundation to design novel point-of-care (POC) diagnostic devices to detect and characterize PMVs in clinical samples.

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Controlling orientation, conformation, and biorecognition of proteins on silane monolayers, conjugate polymers, and thermo-responsive polymer brushes: investigations using TOF-SIMS and principal component analysis

2020

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Control over orientation and conformation of surface-immobilized proteins, determining their biological activity, plays a critical role in biointerface engineering. Specific protein state can be achieved with adjusted surface preparation and immobilization conditions through different types of protein-surface and protein-protein interactions, as outlined in this work. Time-of-flight secondary ion mass spectroscopy, combining surface sensitivity with excellent chemical specificity enhanced by multivariate data analysis, is the most suited surface analysis method to provide information about protein state. This work highlights recent applications of the multivariate principal component analysis of TOF-SIMS spectra to trace orientation and conformation changes of various proteins (antibody, bovine serum albumin, and streptavidin) immobilized by adsorption, specific binding, and covalent attachment on different surfaces, including self-assembled monolayers on silicon, solution-deposited polythiophenes, and thermo-responsive polymer brushes. Multivariate TOF-SIMS results correlate well with AFM data and binding assays for antibody-antigen and streptavidin-biotin recognition. Additionally, several novel extensions of the multivariate TOF-SIMS method are discussed.

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Indirect immunoassay on functionalized silicon surface: Molecular arrangement, composition and orientation examined step-by-step with multi-technique and multivariate analysis

Cited by 14 publications

References 32 publications

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

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

Using a lactadherin-immobilized silicon surface for capturing and monitoring plasma microvesicles as a foundation for diagnostic device development

Controlling orientation, conformation, and biorecognition of proteins on silane monolayers, conjugate polymers, and thermo-responsive polymer brushes: investigations using TOF-SIMS and principal component analysis

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