In this paper we report an acid-modulated strategy for novel peptide microarray production on biosensor interfaces. We initially selected a controlled pore glass (CPG) as a support for solid-phase peptide synthesis (SPPS) to implement a chemistry that can be performed at the interface of multiple field effect transistor (FET) sensors, eventually to generate label-free peptide microarrays for protein screening. Our chemistry uses a temporary protection of the N-terminal amino function of each amino acid building block with a tert-butyloxycarbonyl (Boc) group that can be removed after each SPPS cycle, in combination with semi-permanent protection of the side chains of trifunctional amino acid residues.
View Article and Find Full Text PDFThe control of acidity drives the assembly of biopolymers that are essential for a wide range of applications. Its miniaturization can increase the speed and the possibilities of combinatorial throughput for their manipulation, similar to the way that the miniaturization of transistors allows logical operations in microelectronics with a high throughput. Here, we present a device containing multiplexed microreactors, each one enabling independent electrochemical control of acidity in ∼2.
View Article and Find Full Text PDFControlling locally produced acidity in miniaturized spaces is of high importance to manage simultaneous chemical reactions. Here, we present a platform that hosts miniaturized micro-reactors each one enabling electrochemical control of the acidity in ∼nL volumes. We demonstrated the local control of chemical reactions with the deprotection of strong acid labile groups in a region of 150 μm of diameter of upstanding glass using high proton concentrations (∼10 M) and the acidity contrasts between the cell region and the outside.
View Article and Find Full Text PDFLabel-free field-effect transistor-based immunosensors are promising candidates for proteomics and peptidomics-based diagnostics and therapeutics due to their high multiplexing capability, fast response time, and ability to increase the sensor sensitivity due to the short length of peptides. In this work, planar junctionless field-effect transistor sensors (FETs) were fabricated and characterized for pH sensing. The device with SiO gate oxide has shown voltage sensitivity of 41.
View Article and Find Full Text PDFIn this work, we present a comprehensive analytical model and results for an absolute pH sensor. Our work aims to address critical scientific issues such as: (1) the impact of the oxide degradation (sensing interface deterioration) on the sensor's performance and (2) how to achieve a measurement of the absolute ion activity. The methods described here are based on analytical equations which we have derived and implemented in MATLAB code to execute the numerical experiments.
View Article and Find Full Text PDFExpert Rev Proteomics
April 2020
Introduction: The importance of biomarkers for pharmaceutical drug development and clinical diagnostics is more significant than ever in the current shift toward personalized medicine. Biomarkers have taken a central position either as companion markers to support drug development and patient selection, or as indicators aiming to detect the earliest perturbations indicative of disease, minimizing therapeutic intervention or even enabling disease reversal. Protein biomarkers are of particular interest given their central role in biochemical pathways.
View Article and Find Full Text PDFWe present a review of field effect transistors (FET) from the point of view of their applications to label-free sensing in the era of genomics and proteomics. Here, rather than a collection of Bio-FET achievements, we propose an analysis of the different issues hampering the use of these devices into clinical applications. We make a particular emphasis on the influence of the sensor geometry in the phenomena of mass transport of analytes, which is a topic that has been traditionally overlooked in the analysis and design of biosensors, but that plays a central role in the achievement of low limits of detection.
View Article and Find Full Text PDFThe development of next generation medicines demands more sensitive and reliable label-free sensing able to cope with increasing needs of multiplexing and shorter times to results. Field effect transistor-based biosensors emerge as one of the main possible technologies to cover the existing gap. The general trend for the sensors has been miniaturization with the expectation of improving sensitivity and response time but presenting issues with reproducibility and noise level.
View Article and Find Full Text PDFTemperature sensing is one of the important features of Micro Electro Mechanical Systems and a monolithic integration provides advantages for both fabrication simplicity and performance. The use of Silicon On Insulator substrates allows simple fabrication of integrated wires that can be used as thermistors. We fabricated rectangular and triangular silicon wires with different dimensions in a single step fabrication process based on the wet etching of a <110> Silicon On Insulator substrate.
View Article and Find Full Text PDFThe electrochemical management of the proton concentration in miniaturized dimensions opens the way to control and parallelize multistep chemical reactions, but still it faces many challenges linked to the efficient proton generation and control of their diffusion. Here we present a device operated electrochemically that demonstrates the control of the pH in a cell of ∼140 nL. The device comprises a microfluidic reactor integrated with a pneumatic mechanism that allows the exchange of reagents and the isolation of protons to decrease the effect of their diffusion.
View Article and Find Full Text PDFIn the following article an electron/ion microscopy study will be presented which investigates the uptake of silver nanoparticles (AgNPs) by the marine diatom Thalassiosira pseudonana, a primary producer aquatic species. This organism has a characteristic silica exoskeleton that may represent a barrier for the uptake of some chemical pollutants, including nanoparticles (NPs), but that presents a technical challenge when attempting to use electron-microscopy (EM) methods to study NP uptake. Here we present a convenient method to detect the NPs interacting with the diatom cell.
View Article and Find Full Text PDFWe report here an in vitro evaluation of silica nanoparticle uptake by lung epithelial cells (A549), the cytotoxic effect of the particles and we propose autophagy as possible survival strategy. The effect of surface charge, serum proteins and the influence of inhibitors on the uptake of 20 nm monodispersed nanoparticles with various functional groups are discussed. Uptake rate of the particles with various functional groups is demonstrated to be similar in the presence of serum proteins, while the uptake rate ranking is COOH>NH2>OH under serum free conditions.
View Article and Find Full Text PDFGold nanoparticles (AuNPs) are currently used in several fields including biomedical applications, although no conclusive information on their cytotoxicity is available. For this reason this work has investigated the effects of AuNPs in vitro on Balb/3T3 mouse fibroblasts. Results obtained exposing cells for 72 h to AuNPs 5 and 15 nm citrate stabilized, revealed cytotoxic effects only for AuNPs 5 nm at concentration ≥ 50 μM if measured by colony forming efficiency (CFE).
View Article and Find Full Text PDFInterleukin 1 beta (IL-1β)-dependent inflammatory disorders, such as rheumatoid arthritis and psoriasis, pose a serious medical burden worldwide, where patients face a lifetime of illness and treatment. Organogold compounds have been used since the 1930s to treat rheumatic and other IL-1β-dependent diseases and, though their mechanisms of action are still unclear, there is evidence that gold interferes with the transmission of inflammatory signalling. Here we show for the first time that citrate-stabilized gold nanoparticles, in a size dependent manner, specifically downregulate cellular responses induced by IL-1β both in vitro and in vivo.
View Article and Find Full Text PDFWe present a novel approach to tackle the most common drawback of using surface plasmon resonance for analyte screening in complex biological matrices--the nonspecific binding to the sensor chip surface. By using a perforated membrane supported by a polymeric gel structure at the evanescent wave penetration depth, we have fabricated a non-fouling sieve above the sensing region. The sieve shields the evanescent wave from nonspecific interactions which interfere with SPR sensing by minimizing the fouled area of the polymeric gel and preventing the translocation of large particles, e.
View Article and Find Full Text PDFIn this work we investigated the toxicological effects of nude and chemically functionalised (-NH(2), -OH and -COOH groups) multiwall carbon nanotubes (mwCNTs) using immortalised mouse fibroblasts cell line (Balb/3T3) as in vitro model, alternative to the use of animals, to assess basal cytotoxicity, carcinogenic potential, genotoxicity and cell interaction of nanomaterials (NM). Combining in vitro tests such as cell transformation assay and micronucleus with physicochemical and topological analysis, we obtained results showing no cytotoxicity and genotoxicity. Carcinogenic potential and mwCNTs interaction with cells were instead evident.
View Article and Find Full Text PDFWe show that using asymmetric flow-field-flow fractionation and UV-vis detector it is possible to separate, characterize, and quantify the correct number size distribution of gold nanoparticle (AuNP) mixtures of various sizes in the 5-60 nm range for which simple dynamic light scattering measurements give misleading information. The size of the collected nanoparticles fractions can be determined both in solution and in the solid state, and their surface chemistry characterized by NMR. This method will find widespread applications both in the process of "size purification" after the synthesis of AuNP and in the identification and characterization of gold-based nanomaterials in consumer products.
View Article and Find Full Text PDFOptical control of the lateral quantum confinement and number of electrons confined in nanofabricated GaAs/AlGaAs quantum dots is achieved by illumination with a weak laser beam that is absorbed in the AlGaAs barrier. Precise tuning of energy-level structure and electron population is demonstrated by monitoring the low-lying transitions of the electrons from the lowest quantum-dot energy shells by resonant inelastic light scattering. These findings open the way to the manipulation of single electrons in these quantum dots without the need of external metallic gates.
View Article and Find Full Text PDFWe report inelastic light scattering measurements of spin and charge excitations in nanofabricated AlGaAs/GaAs quantum dots with few electrons. A narrow spin excitation peak is observed and assigned to the intershell triplet-to-singlet monopole mode of dots with four electrons. Configuration-interaction theory provides precise quantitative interpretations that uncover large correlation effects that are comparable to exchange Coulomb interactions.
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