Surface Preparation Strategies for Improved Parallelization and Reproducible MALDI-TOF MS Ligand Binding Assays

Roth, Michael J.; Maresh, Erica M.; Plymire, Daniel A.; Zhang, Junmei; Corbett, J. R.; Robbins, R.O.; Patrie, Steven M.

doi:10.1021/am3024756

Cited by 11 publications

(6 citation statements)

References 23 publications

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“…A multitude of approaches to minimize background interference while maximizing compound ionization have been reported, suggesting a variety of matrix constituents, additives, solid supports, and preparation procedures. [22][23][24][25][26] Many of these improvements, however, are valid only for specific analyte chemotypes, require special treatment of MALDI target plates, or interfere with automation processes. Therefore, a set of 10 different MALDI matrices, which have proven to be compatible with the applied automated spotting procedure, were evaluated for suitability.…”

Section: Optimization Of Maldi Matrix Composition and Deposition For mentioning

confidence: 99%

MALDI-TOF-Based Affinity Selection Mass Spectrometry for Automated Screening of Protein–Ligand Interactions at High Throughput

et al. 2021

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Section: Optimization Of Maldi Matrix Composition and Deposition For mentioning

confidence: 99%

MALDI-TOF-Based Affinity Selection Mass Spectrometry for Automated Screening of Protein–Ligand Interactions at High Throughput

et al. 2021

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“…TLT/ VIC on arrays was performed on an apparatus previously described with MALDI matrix α-cyano-4-hydroxycinnamic acid (CHCA) sublimed at a ∼25 ng/mm 2 surface density and VIC performed with dimethylformamide (Figure S-1). 24,42 Surface ADAMTS-13 Activity Assays. Ten microliters of plasma was diluted in 50 mM MES buffer with 20 mM CaCl 2 at pH 6.25.…”

Section: ■ Methodsmentioning

confidence: 99%

Measurement of Blood Protease Kinetic Parameters with Self-Assembled Monolayer Ligand Binding Assays and Label-Free MALDI-TOF MS

et al. 2013

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We report novel ligand binding assay (LBA) surface modalities that permit plasma protease catalytic efficiency (kcat/km) determination by MALDI-TOF MS without the use of liquid chromatography or internal standards such as chemical or metalized labels. Two model LBAs were constructed on planar self-assembled monolayers (SAMs) and used to evaluate the clinically relevant metalloprotease ADAMTS-13 kinetics in plasma. The SAM chemistries were designed to improve biosampling efficiency by minimization of nonspecific adsorption of abundant proteins present at ~100,000× the concentration of the endogenous enzyme. In the first protocol, in-solution digestion of the ADAMTS-13 substrate (vWFh) was performed with immunoaffinity enrichment of the reaction substrate and product to SAM arrays. The second configuration examined protease kcat/km via a surface digestion modality where different substrates were covalently immobilized to the SAM at controlled surface density for optimized protease screens. The results show the MALDI-TOF MS LBA platforms provide limits of quantitation to ~1% protease activity (~60 pM enzyme concentration) in <1 h analysis time, a ~16× improvement over other MS-based LBA formats. Implementation of a vacuum-sublimed MALDI matrix provided good MALDI-TOF MS intra- and interday repeatability, ~1.2 and ~6.6% RSD, respectively. Platform reliability permitted kcat/km determination without internal standards with observed values ~10× improved versus conventional fluorophoric assays. Application of the assays to 12 clinical plasma samples demonstrated proof-of-concept for clinical applications. Overall, this work demonstrates that rationally designed surface chemistries for MALDI-TOF MS may serve as an alternative, label-free methodology with potential for a wide range of biotechnology applications related to targeted enzyme molecular diagnostics.

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“…Protein G and Protein A have been used as IgG capture molecules in immunosensor devices fabricated using self-assembled monolayers (Makaraviciute and Ramanaviciene 2013 ; Trilling et al 2013 ). The IgG capture molecules have been immobilised to surfaces using self-assembled monolayers of thiolalkanes functionalised with carboxylic acid groups for amine cross-linking (Jyoung et al 2006 ), nitrolotriacetic acid (NTA) for binding proteins with a hexa-histidine tag (Roth et al 2013 ), or for binding biotinlyated proteins (Jung et al 2006 ). Monolayers of proteins A and G have been formed by direct assembly onto gold by incorporating thiol groups (Oh et al 2004 ), or through the use of fusion proteins such as glutathione S -transferase (GST) (Ha et al 2006 ) or elastin (Gao et al 2006 ).…”

Section: Introductionmentioning

confidence: 99%

Engineered self-assembling monolayers for label free detection of influenza nucleoprotein

Soliakov

Shah

Holt

et al. 2015

Biomed Microdevices

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Integrating nanotechnology into useable devices requires a combination of bottom up and top down methodology. Often the techniques to measure and control these different components are entirely different, so methods that can analyse the nanoscale component in situ are of increasing importance. Here we describe a strategy that employs a self-assembling monolayer of engineered protein chimeras to display an array of oriented antibodies (IgG) on a microelectronic device for the label free detection of influenza nucleoprotein. The structural and functional properties of the bio-interface were characterised by a range of physical techniques including surface plasmon resonance, quartz-crystal microbalance and neutron reflectometry. This combination of methods reveals a 13.5 nm thick engineered-monolayer that (i) self-assembles on gold surfaces, (ii) captures IgG with high affinity in a defined orientation and (iii) specifically recognises the influenza A nucleoprotein. Furthermore we also show that this non-covalent self-assembled structure can render the dissociation of bound IgG irreversible by chemical crosslinking in situ without affecting the IgG function. The methods can thus describe in detail the transition from soluble engineered molecules with nanometre dimensions to an array that demonstrates the principles of a working influenza sensor.Electronic supplementary materialThe online version of this article (doi:10.1007/s10544-015-9951-z) contains supplementary material, which is available to authorized users.

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Surface Preparation Strategies for Improved Parallelization and Reproducible MALDI-TOF MS Ligand Binding Assays

Cited by 11 publications

References 23 publications

MALDI-TOF-Based Affinity Selection Mass Spectrometry for Automated Screening of Protein–Ligand Interactions at High Throughput

MALDI-TOF-Based Affinity Selection Mass Spectrometry for Automated Screening of Protein–Ligand Interactions at High Throughput

Measurement of Blood Protease Kinetic Parameters with Self-Assembled Monolayer Ligand Binding Assays and Label-Free MALDI-TOF MS

Engineered self-assembling monolayers for label free detection of influenza nucleoprotein

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