Anne-Laure Gassner

Soft Stylus Probes for Scanning Electrochemical Microscopy

A soft stylus microelectrode probe has been developed to carry out scanning electrochemical microscopy (SECM) of rough, tilted, and large substrates in contact mode. It is fabricated by first ablating a microchannel in a polyethylene terephthalate thin film and filling it with a conductive carbon ink. After curing the carbon track and lamination with a polymer film, the V-shaped stylus was cut thereby forming a probe, with the cross section of the carbon track at the tip being exposed either by UV-photoablation machining or by blade cutting followed by polishing to produce a crescent moon-shaped carbon microelectrode. The probe properties have been assessed by cyclic voltammetry, approach curves, and line scans over electrochemically active and inactive substrates of different roughness. The influence of probe bending on contact mode imaging was then characterized using simple patterns. Boundary element method simulations were employed to rationalize the distance-dependent electrochemical response of the soft stylus probes.

Capillary electrophoresis immunoassay using magnetic beads

Protein A‐coated magnetic beads (0.3 μm) have been trapped in a small portion of a neutrally coated capillary (50 μm id). Anti‐β‐lactoglobulin (β‐LG) antibodies have then been immobilized on the beads through strong affinity with protein A to subsequently capture β‐LG from model or real samples. Once the immunocomplexes formed at physiological pH, a discontinuous buffer system has been used to release the partners and preconcentrate them by transient ITP. The antigens and antibodies have finally been separated by CZE and detected by UV absorbance. An LOQ of 55 nM has been achieved. This methodology has been applied to quantify native β‐LG in pasteurized and ultra‐high‐temperature‐treated bovine milk. All the described procedures, including immunosorbent preparation, sample extraction, cleanup, preconcentration, and separation are completely automated on a commercial CE instrument. As this CE immunoassay method is simple, rapid, selective, and sensitive, it should be a practical and attractive technology for the analysis of complicated biological samples.

Analysis of major milk whey proteins by immunoaffinity capillary electrophoresis coupled with MALDI-MS

Two major milk whey proteins, β‐lactoglobulin and α‐lactalbumin, are among the main cow milk allergens and can cause allergy even at a very low concentrations. Therefore, these proteins are interesting targets in food analysis, not only for food quality control but also for highlighting the presence of allergens. Herein, a sensitive analysis for β‐lactoglobulin and α‐lactalbumin was developed using immunoaffinity capillary electrophoresis hyphenated with MALDI‐MS. Magnetic beads functionalized with appropriate antibodies were used for β‐lactoglobulin and α‐lactalbumin immunocapture inside the capillary. After elution from the beads, analyte focusing and separation were performed by transient isotachophoresis followed by MALDI‐MS analysis performed through an automated iontophoretic fraction collection interface. A LOD in the low nanomolar range was attained for both whey proteins. The method developed was further applied to the analysis of different milk samples including fortified soy milk.

Ring magnets for magnetic beads trapping in a capillary

This paper introduces the concept of ring magnets for magnetic beads (MBs) trapping in a capillary. Such magnets enable an easy insertion of a capillary simply like a pearl on a string. With this system, high magnetic forces are obtained thanks to the proximity between the magnet and the capillary, giving the opportunity to work at higher flow rates than with classical setups using two magnets with their magnetization perpendicular to the capillary. Moreover, by alternating magnets and non-magnetic spacers either in attraction or repulsion configuration, it is possible to form a chain and as a consequence to adapt the number of magnets to the desired number of plugs, thus controlling the surface available for molecule binding. Magnetic force mapping was first carried out by numerical simulations for a single ring magnet. The usefulness of this concept was then demonstrated with the achievement of an immunoassay and an online preconcentration experiment. To study the formation of multiplugs, the magnetic force was first simulated for a chain of four magnets in repulsion. This force was then introduced into a convection-diffusion model to understand the influence of the flow velocity on their size and position. The numerical simulations were qualitatively corroborated by microscopic visualizations, carried out in a capillary placed between rectangular magnets having a magnetization parallel to the capillary, and quantitatively by bead capture efficiency experiments.

Bubble cell for magnetic bead trapping in capillary electrophoresis

AbstractA bubble cell capillary classically used to extend the optical path length for UV–vis detection is employed here to trap magnetic beads. With this system, a large amount of beads can be captured without inducing a strong pressure drop, as it is the case with magnetic beads trapped in a standard capillary, thereby having less effect on the experimental conditions. Using numerical simulations and microscopic visualizations, the capture of beads inside a bubble cell was investigated with two magnet configurations. Pressure-driven and electro-osmotic flow velocities were measured for different amounts of protein-A-coated beads or C18-functionalized beads (RPC-18). Solid-phase extraction of a model antibody on protein-A beads and preconcentration of fluorescein on RPC-18 beads were performed as proof of concept experiments. FigureIsovalues of the magnetic induction produced by two permanent magnets in attraction configuration with a capillary placed between them.