7 research outputs found
Biochip Spray: Simplified Coupling of Surface Plasmon Resonance Biosensing and Mass Spectrometry
Multiplex surface plasmon resonance biosensing and its transferability towards imaging nanoplasmonics for detection of mycotoxins in barley
A 6-plex competitive inhibition immunoassay for mycotoxins in barley was developed on a prototype portable nanostructured imaging surface plasmon resonance (iSPR) instrument, also referred to as imaging nanoplasmonics. As a benchmark for the prototype nanoplasmonics instrument, first a double 3-plex assay was developed for the detection of deoxynivalenol (DON), zearalenone (ZEA), T-2 toxin (T-2), ochratoxin A (OTA), fumonisin B1 (FB1) and aflatoxin B1 (AFB1) using a well-established benchtop SPR instrument and two biosensor chips. To this end, ovalbumin (OVA) conjugates of mycotoxins were immobilized on the chip via amine coupling. The SPR response was then recorded upon injection of a mixture of antibodies at a fixed concentration and the sample (or matrix-matched standard) over a chip with immobilized mycotoxin-OVA conjugates. The chips were regenerated after each sample using 10 mM HCl and 20 mM NaOH and could be used for at least 60 cycles. The limits of detection in barley (in μg kg-1) were determined to be 26 for DON, 6 for ZEA, 0.6 for T-2, 3 for OTA, 2 for FB1 and 0.6 for AFB1. Preliminary in-house validation showed that DON, T-2, ZEA and FB1 can be detected at the European Union regulatory limits, while for OTA and AFB1 sensitivities should be improved. Furthermore, measurement of naturally contaminated barley showed that the assay can be used as a semi-quantitative screening method for mycotoxins prior to liquid chromatography tandem mass spectrometry (LC-MS/MS). Finally, using the same bio-reagents the assay was transferred to a 6-plex format in the nanoplasmonics instrument and subsequently the two assays were compared. Although less sensitive, the 6-plex portable iSPR assay still allowed detection of DON, T-2, ZEA and FB1 at relevant levels. Therefore, the prototype iSPR shows potential for future applications in rapid in-field and at-line screening of multiple mycotoxins.</p
Analysis of Mycotoxins in Beer Using a Portable Nanostructured Imaging Surface Plasmon Resonance Biosensor
A competitive inhibition immunoassay is described for the mycotoxins deoxynivalenol (DON) and ochratoxin A (OTA) in beer using a portable nanostructured imaging surface plasmon resonance (iSPR) biosensor, also referred to as imaging nanoplasmonics. The toxins were directly and covalently immobilized on a 3-dimensional carboxymethylated dextran (CMD) layer on a nanostructured iSPR chip. The assay is based on competition between the immobilized mycotoxins and free mycotoxins in the solution for binding to specific antibodies. The chip surface was regenerated after each cycle, and the combination of CMD and direct immobilization of toxins allowed the chips to be used for more than 450 cycles. The limits of detection (LODs) in beer were 17 ng/mL for DON and 7 ng/mL for OTA (or 0.09 ng/mL after 75 times enrichment). These LODs allowed detection of even less than 10% depletion of the tolerable daily intake of DON and OTA by beer. Significant cross-reactivity of anti-DON was observed toward DON-3-glucoside and 3-acetyl-DON, while no cross-reactivity was seen for 15-acetyl-DON. A preliminary in-house validation with 20 different batches of beer showed that both toxins can be detected at the considered theoretical safe level for beer. The assay can be used for in-field or at-line detection of DON in beer and also in barley without preconcentration, while OTA in beer requires an additional enrichment step, thus making the latter in its present form less suitable for field applications
Biochip Spray: Simplified Coupling of Surface Plasmon Resonance Biosensing and Mass Spectrometry
A simplified coupling
of surface plasmon resonance (SPR) immuno-biosensing
with ambient ionization mass spectrometry (MS) was developed. It combines
two orthogonal analysis techniques: the biosensing capability of SPR
and the chemical identification power of high resolution MS. As a
proof-of-principle, deoxynivalenol (DON), an important mycotoxin,
was captured using an SPR gold chip containing an antifouling layer
and monoclonal antibodies against the toxin and, after washing, the
chip could be taken out and analyzed by direct spray MS of the biosensor
chip to confirm the identity of DON. Furthermore, cross-reacting conjugates
of DON present in a naturally contaminated beer could be successfully
identified, thus showing the potential of rapid identification of
(un)Âexpected cross-reacting molecules
Ambient Surface Analysis of Organic Monolayers using Direct Analysis in Real Time Orbitrap Mass Spectrometry
A better characterization of nanometer-thick
organic layers (monolayers)
as used for engineering surface properties, biosensing, nanomedicine,
and smart materials will widen their application. The aim of this
study was to develop direct analysis in real time high-resolution
mass spectrometry (DART-HRMS) into a new and complementary analytical
tool for characterizing organic monolayers. To assess the scope and
formulate general interpretation rules, DART-HRMS was used to analyze
a diverse set of monolayers having different chemistries (amides,
esters, amines, acids, alcohols, alkanes, ethers, thioethers, polymers,
sugars) on five different substrates (Si, Si<sub>3</sub>N<sub>4</sub>, glass, Al<sub>2</sub>O<sub>3</sub>, Au). The substrate did not
play a major role except in the case of gold, for which breaking of
the weak Au–S bond that tethers the monolayer to the surface,
was observed. For monolayers with stronger covalent interfacial bonds,
fragmentation around terminal groups was found. For ester and amide-terminated
monolayers, in situ hydrolysis during DART resulted in the detection
of ions characteristic of the terminal groups (alcohol, amine, carboxylic
acid). For ether and thioether-terminated layers, scission of C–O
or C–S bonds also led to the release of the terminal part of
the monolayer in a predictable manner. Only the spectra of alkane
monolayers could not be interpreted. DART-HRMS allowed for the analysis
of and distinction between monolayers containing biologically relevant
mono or disaccharides. Overall, DART-HRMS is a promising surface analysis
technique that combines detailed structural information on nanomaterials
and ultrathin films with fast analyses under ambient conditions
Ambient Surface Analysis of Organic Monolayers using Direct Analysis in Real Time Orbitrap Mass Spectrometry
A better characterization of nanometer-thick
organic layers (monolayers)
as used for engineering surface properties, biosensing, nanomedicine,
and smart materials will widen their application. The aim of this
study was to develop direct analysis in real time high-resolution
mass spectrometry (DART-HRMS) into a new and complementary analytical
tool for characterizing organic monolayers. To assess the scope and
formulate general interpretation rules, DART-HRMS was used to analyze
a diverse set of monolayers having different chemistries (amides,
esters, amines, acids, alcohols, alkanes, ethers, thioethers, polymers,
sugars) on five different substrates (Si, Si<sub>3</sub>N<sub>4</sub>, glass, Al<sub>2</sub>O<sub>3</sub>, Au). The substrate did not
play a major role except in the case of gold, for which breaking of
the weak Au–S bond that tethers the monolayer to the surface,
was observed. For monolayers with stronger covalent interfacial bonds,
fragmentation around terminal groups was found. For ester and amide-terminated
monolayers, in situ hydrolysis during DART resulted in the detection
of ions characteristic of the terminal groups (alcohol, amine, carboxylic
acid). For ether and thioether-terminated layers, scission of C–O
or C–S bonds also led to the release of the terminal part of
the monolayer in a predictable manner. Only the spectra of alkane
monolayers could not be interpreted. DART-HRMS allowed for the analysis
of and distinction between monolayers containing biologically relevant
mono or disaccharides. Overall, DART-HRMS is a promising surface analysis
technique that combines detailed structural information on nanomaterials
and ultrathin films with fast analyses under ambient conditions