On-line Coupling of Aptamer Affinity Solid-Phase Extraction and Immobilized Enzyme Microreactor Capillary Electrophoresis-Mass Spectrometry for the Sensitive Targeted Bottom-Up Analysis of Protein Biomarkers.

In this paper, we present a fully integrated valve-free method for the sensitive targeted bottom-up analysis of proteins through on-line aptamer affinity solid-phase extraction and immobilized enzyme microreactor capillary electrophoresis-mass spectrometry (AA-SPE-IMER-CE-MS). The method was developed analyzing α-synuclein (α-syn), which is a protein biomarker related to different neurodegenerative disorders, including Parkinson's disease. Under optimized conditions, on-line purification and preconcentration of α-syn, enzymatic digestion, electrophoretic separation, and identification of the tryptic peptides by mass spectrometry was achieved in less than 35 min. The limit of detection was 0.02 µg mL-1 of digested protein (66.7% of coverage, i.e., 8 out of 12 expected tryptic peptides were detected). This value was 125 and 10 times lower than for independent on-line digestion by IMER-CE-MS (2.5 µg mL-1) and on-line preconcentration by AA-SPE-CE-MS (0.2 µg mL-1). The repeatability of AA-SPE-IMER-CE-MS was adequate (at 0.5 µg mL-1,% RSD ranged from 3.7 to 16.9% for peak areas and 3.5 to 7.7% for migration times of the tryptic peptides), and the modified capillary could be reused up to 10 analyses with optimum performance, similarly to IMER-CE-MS. The method was subsequently applied to the analysis of endogenous α-syn from red blood cell lysates. Ten α-syn tryptic peptides were detected (83.3% of coverage), enabling the characterization and localization of post-translational modifications of blood α-syn (i.e., N-terminal acetylation).

Nanomaterials-Based Sensors for Respiratory Viral Detection: A Review.

Contagious diseases are the principal cause of mortality, particularly respiratory viruses, a real menace for public health and economic development worldwide. Therefore, timely diagnosis and treatments are the only life-saving strategy to overcome any epidemic and particularly the ongoing prevailing pandemic COVID-19 caused by SARS-CoV-2. A rapid identification, point of care, portable, highly sensitive, stable, and inexpensive device is needed which is exceptionally satisfied by sensor technology. Consequently, the researchers have directed their attention to employing sensors targeting multiple analyses of pathogenic detections across the world. Nanostructured materials (nanoparticles, nanowires, nanobundles, etc.), owing to their unique characteristics such as large surface-to-volume ratio and nanoscale interactions, are widely employed to fabricate facile sensors to meet all the immediate emerging challenges and threats. This review is anticipated to foster researchers in developing advanced nanomaterials-based sensors for the increasing number of COVID-19 cases across the globe. The mechanism of respiratory viral detection by nanomaterials-based sensors has been reported. Moreover, the advantages, disadvantages, and their comparison with conventional sensors are summarized. Furthermore, we have highlighted the challenges and future potential of these sensors for achieving efficient and rapid detection.

On-line aptamer affinity solid-phase extraction direct mass spectrometry for the rapid analysis of α-synuclein in blood.

On-line aptamer affinity solid-phase extraction direct mass spectrometry (AA-SPE-MS) is presented for the rapid purification, preconcentration, and characterization of α-synuclein (α-syn), which is a protein biomarker related to Parkinson's disease. Valve-free AA-SPE-MS is easily implemented using the typical SPE microcartridges and instrumental set-up necessary for on-line aptamer affinity solid-phase extraction capillary electrophoresis-mass spectrometry (AA-SPE-CE-MS). The essential requirement is substituting the application of the separation voltage by a pressure of 100 mbar for mobilization of the eluted protein through the capillary towards the mass spectrometer. Under optimized conditions with recombinant α-syn, repeatability is good in terms of migration time and peak area (percent relative standard deviation (%RSD) values (n = 3) are 1.3 and 6.6% at 1 µg mL-1, respectively). The method is satisfactorily linear between 0.025 and 5 µg mL-1 (R2 > 0.986), and limit of detection (LOD) is 0.02 µg mL-1 (i.e. 1000, 500, and 10 times lower than by CE-MS, direct MS, and AA-SPE-CE-MS, respectively). The established AA-SPE-MS method is further compared with AA-SPE-CE-MS, including for the analysis of α-syn in blood. The comparison discloses the advantages and disadvantages of AA-SPE-MS for the rapid and sensitive targeted analysis of protein biomarkers in biological fluids.

Nanomaterials-based sensors for the detection of COVID-19: A review.

With the threat of increasing SARS-CoV-2 cases looming in front of us and no effective and safest vaccine available to curb this pandemic disease due to its sprouting variants, many countries have undergone a lockdown 2.0 or planning a lockdown 3.0. This has upstretched an unprecedented demand to develop rapid, sensitive, and highly selective diagnostic devices that can quickly detect coronavirus (COVID-19). Traditional techniques like polymerase chain reaction have proven to be time-inefficient, expensive, labor intensive, and impracticable in remote settings. This shifts the attention to alternative biosensing devices that can be successfully used to sense the COVID-19 infection and curb the spread of coronavirus cases. Among these, nanomaterial-based biosensors hold immense potential for rapid coronavirus detection because of their noninvasive and susceptible, as well as selective properties that have the potential to give real-time results at an economical cost. These diagnostic devices can be used for mass COVID-19 detection to understand the rapid progression of the infection and give better-suited therapies. This review provides an overview of existing and potential nanomaterial-based biosensors that can be used for rapid SARS-CoV-2 diagnostics. Novel biosensors employing different detection mechanisms are also highlighted in different sections of this review. Practical tools and techniques required to develop such biosensors to make them reliable and portable have also been discussed in the article. Finally, the review is concluded by presenting the current challenges and future perspectives of nanomaterial-based biosensors in SARS-CoV-2 diagnostics.

Fourth-generation glucose sensors composed of copper nanostructures for diabetes management: A critical review.

More than five decades have been invested in understanding glucose biosensors. Yet, this immensely versatile field has continued to gain attention from the scientific world to better understand and diagnose diabetes. However, such extensive work done to improve glucose sensing devices has still not yielded desirable results. Drawbacks like the necessity of the invasive finger-pricking step and the lack of optimization of diagnostic interventions still need to be considered to improve the testing process of diabetic patients. To upgrade the glucose-sensing devices and reduce the number of intermediary steps during glucose measurement, fourth-generation glucose sensors (FGGS) have been introduced. These sensors, made using robust electrocatalytic copper nanostructures, improve diagnostic efficiency and cost-effectiveness. This review aims to present the essential scientific progress in copper nanostructure-based FGGS in the past 10 years (2010 to present). After a short introduction, we presented the working principles of these sensors. We then highlighted the importance of copper nanostructures as advanced electrode materials to develop reliable real-time FGGS. Finally, we cover the advantages, shortcomings, and prospects for developing highly sensitive, stable, and specific FGGS.

A simple method for the analysis of extracellular vesicles enriched for exosomes from human serum by capillary electrophoresis with ultraviolet diode array detection.

Extracellular vesicles (EVs) are membrane enclosed vesicles (<1 µm), such as exosomes (30-150 nm), involved in cell communication, which have important biological implications. In this study, EV preparations were enriched for exosomes from human serum by polyethylene glycol (PEG) precipitation. Different variables of the PEG precipitation method (i.e. concentration of PEG, filtration and centrifugation of the resuspended pellets) were evaluated by measuring the size of the isolated particles by dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA). In addition, a novel capillary electrophoresis-ultraviolet diode array (CE-UV-DAD) method was developed to obtain characteristic multiwavelength electrophoretic profiles of the EV preparations. Using EV preparations precipitated with 10% m/v of PEG, a background electrolyte (BGE) of 0.1 M Tris and 0.25 M boric acid at pH 7.9 with 0.5% m/v of hydroxypropyl cellulose (HPC) allowed reducing the adsorption of the EVs to the inner wall of the fused silica separation capillary. Sodium dodecyl sulfate (SDS) at 0.1% m/v was also necessary to enhance dispersibility, while homogenizing the charge of the particles to improve the size-dependent separation induced by HPC. Under these optimized conditions, a characteristic electrophoretic multiwavelength profile of the EV preparation and a standard of exosomes was obtained, and separation showed excellent reproducibility and appropriate analysis times. The obtained electrophoretic fingerprints are a simple, effective and complementary tool for the quality control of EV preparations.

Recent Advances in Non-Enzymatic Glucose Sensors Based on Metal and Metal Oxide Nanostructures for Diabetes Management- A Review.

There is an undeniable growing number of diabetes cases worldwide that have received widespread global attention by many pharmaceutical and clinical industries to develop better functioning glucose sensing devices. This has called for an unprecedented demand to develop highly efficient, stable, selective, and sensitive non-enzymatic glucose sensors (NEGS). Interestingly, many novel materials have shown the promising potential of directly detecting glucose in the blood and fluids. This review exclusively encompasses the electrochemical detection of glucose and its mechanism based on various metal-based materials such as cobalt (Co), nickel (Ni), zinc (Zn), copper (Cu), iron (Fe), manganese (Mn), titanium (Ti), iridium (Ir), and rhodium (Rh). Multiple aspects of these metals and their oxides were explored vis-à-vis their performance in glucose detection. The direct glucose oxidation via metallic redox centres is explained by the chemisorption model and the incipient hydrous oxide/adatom mediator (IHOAM) model. The glucose electrooxidation reactions on the electrode surface were elucidated by equations. Furthermore, it was explored that an effective detection of glucose depends on the aspect ratio, surface morphology, active sites, structures, and catalytic activity of nanomaterials, which plays an indispensable role in designing efficient NEGS. The challenges and possible solutions for advancing NEGS have been summarized.

Ionic matrices for matrix-assisted laser desorption/ionization mass spectrometry analysis of microRNA biomarkers.

The use of ionic matrices (IMs) was evaluated as an alternative to conventional matrices to analyze microRNAs (miRNAs) by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 2, 4, 6-Trihydroxyacetophenone (THAP), 6-aza-2-thiothymine (ATT) and 3-hydroxypicolinic acid (3-HPA) and their IMs with pyridine (PYR) and butylamine (BA) were studied to analyze a standard mixture of miRNAs: miR-21, let-7g and iso-miR-16. Among all the studied matrices, ATT-PYR at 75 mg/mL in acetonitrile (MeCN):H2O (50:50, v/v) was selected as the optimal. Furthermore, addition of ammonium citrate dibasic (AC) as signal enhancer was mandatory to obtain an appropriate miRNA detection. ATT-PYR provided the best sensitivity, with limit of detection (LOD) up to 5 nM (equivalent to 1 fmol in the spot) and excellent spot-to-spot repeatability due to the improved homogeneity of the spots compared to the conventional matrices. The applicability of the established method to direct, multiplex and untargeted analysis of miRNAs in serum samples was also investigated.