scholarly journals Kinetic Exclusion Assay of Biomolecules by Aptamer Capture

Sensors ◽  
2020 ◽  
Vol 20 (12) ◽  
pp. 3442
Author(s):  
Mark H. Smith ◽  
Daniel Fologea

DNA aptamers are short nucleotide oligomers selected to bind a target ligand with affinity and specificity rivaling that of antibodies. These remarkable features recommend aptamers as candidates for analytical and therapeutic applications that traditionally use antibodies as biorecognition elements. Numerous traditional and emerging analytical techniques have been proposed and successfully implemented to utilize aptamers for sensing purposes. In this work, we exploited the analytical capabilities offered by the kinetic exclusion assay technology to measure the affinity of fluorescent aptamers for their thrombin target and quantify the concentration of analyte in solution. Standard binding curves constructed by using equilibrated mixtures of aptamers titrated with thrombin were fitted with a 1:1 binding model and provided an effective Kd of the binding in the sub-nanomolar range. However, our experimental results suggest that this simple model does not satisfactorily describe the binding process; therefore, the possibility that the aptamer is composed of a mixture of two or more distinct Kd populations is discussed. The same standard curves, together with a four-parameter logistic equation, were used to determine “unknown” concentrations of thrombin in mock samples. The ability to identify and characterize complex binding stoichiometry, together with the determination of target analyte concentrations in the pM–nM range, supports the adoption of this technology for kinetics, equilibrium, and analytical purposes by employing aptamers as biorecognition elements.

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1768
Author(s):  
Miroslav Rievaj ◽  
Eva Culková ◽  
Damiána Šandorová ◽  
Zuzana Lukáčová-Chomisteková ◽  
Renata Bellová ◽  
...  

This short review deals with the properties and significance of the determination of selenium, which is in trace amounts an essential element for animals and humans, but toxic at high concentrations. It may cause oxidative stress in cells, which leads to the chronic disease called selenosis. Several analytical techniques have been developed for its detection, but electroanalytical methods are advantageous due to simple sample preparation, speed of analysis and high sensitivity of measurements, especially in the case of stripping voltammetry very low detection limits even in picomoles per liter can be reached. A variety of working electrodes based on mercury, carbon, silver, platinum and gold materials were applied to the analysis of selenium in various samples. Only selenium in oxidation state + IV is electroactive therefore the most of voltammetric determinations are devoted to it. However, it is possible to detect also other forms of selenium by indirect electrochemistry approach.


Amino Acids ◽  
2021 ◽  
Author(s):  
Grażyna Gałęzowska ◽  
Joanna Ratajczyk ◽  
Lidia Wolska

AbstractThe quantitation and qualification of amino acids are most commonly used in clinical and epidemiological studies, and provide an excellent way of monitoring compounds in human fluids which have not been monitored previously, to prevent some diseases. Because of this, it is not surprising that scientific interest in evaluating these compounds has resurfaced in recent years and has precipitated the development of a multitude of new analytical techniques. This review considers recent developments in HPLC analytics on the basis of publications from the last few years. It helps to update and systematize knowledge in this area. Particular attention is paid to the progress of analytical methods, pointing out the advantages and drawbacks of the various techniques used for the preparation, separation and determination of amino acids. Depending on the type of sample, the preparation conditions for HPLC analysis change. For this reason, the review has focused on three types of samples, namely urine, blood and cerebrospinal fluid. Despite time-consuming sample preparation before HPLC analysis, an additional derivatization technique should be used, depending on the detection technique used. There are proposals for columns that are specially modified for amino acid separation without derivatization, but the limit of detection of the substance is less beneficial. In view of the fact that amino acid analyses have been performed for years and new solutions may generate increased costs, it may turn out that older proposals are much more advantageous.


Metals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 736
Author(s):  
Peter Seidel ◽  
Doreen Ebert ◽  
Robert Schinke ◽  
Robert Möckel ◽  
Simone Raatz ◽  
...  

Better quality control for alloy manufacturing and sorting of post-consumer scraps relies heavily on the accurate determination of their chemical composition. In recent decades, analytical techniques, such as X-ray fluorescence spectroscopy (XRF), laser-induced breakdown spectroscopy (LIBS), and spark optical emission spectroscopy (spark-OES), found widespread use in the metal industry, though only a few studies were published about the comparison of these techniques for commercially available alloys. Hence, we conducted a study on the evaluation of four analytical techniques (energy-dispersive XRF, wavelength-dispersive XRF, LIBS, and spark-OES) for the determination of metal sample composition. It focuses on the quantitative analysis of nine commercial alloys, representing the three most important alloy classes: copper, aluminum, and steel. First, spark-OES is proven to serve as a validation technique in the use of certified alloy reference samples. Following an examination of the lateral homogeneity by XRF, the results of the techniques are compared, and reasons for deviations are discussed. Finally, a more general evaluation of each technique with its capabilities and limitations is given, taking operation-relevant parameters, such as measurement speed and calibration effort, into account. This study shall serve as a guide for the routine use of these methods in metal producing and recycling industries.


2003 ◽  
Vol 320 (2) ◽  
pp. 292-298 ◽  
Author(s):  
Maria J Vazquez ◽  
Beatriz Rodriguez ◽  
Cleofe Zapatero ◽  
David G Tew

1992 ◽  
Vol 02 (04) ◽  
pp. 489-491 ◽  
Author(s):  
MOMOKO CHIBA ◽  
VENKATESH G. IYENGAR

Tin (Sn) is one of the causative elements of the environmental pollution. As no certified reference materials for Sn are presently available, existing reference materials were analyzed for Sn by two independent analytical techniques; atomic absorption spectrometry (AAS) and neutron activation analysis (NAA). The results obtained by both methods were in agreement except for mixed diet which contains Sn in the range of 50 μ g/g. Further, tin concentrations in human and animal organs have been examined by AAS. Among organs tested tin concentrations in testes were the highest, 2.08±0.62 μ g/g dry weight (mean ±SD, n=12) in humans, and 1.45±0.55 μ g/g (n=8) in mice.


RSC Advances ◽  
2016 ◽  
Vol 6 (71) ◽  
pp. 66963-66970 ◽  
Author(s):  
Haiguan Yang ◽  
Junqi Zha ◽  
Peng Zhang ◽  
Yuhao Xiong ◽  
Linjing Su ◽  
...  

CoS, which was prepared using a facile solvothermal method, and characterized using various analytical techniques, was demonstrated for the first time to exhibit intrinsic peroxidase-like activity.


Chemosensors ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 72
Author(s):  
Aurelia Magdalena Pisoschi ◽  
Aneta Pop ◽  
Florin Iordache ◽  
Loredana Stanca ◽  
Liviu Bilteanu ◽  
...  

Antioxidants are compounds that prevent or delay the oxidation process, acting at a much smaller concentration, in comparison to that of the preserved substrate. Primary antioxidants act as scavenging or chain breaking antioxidants, delaying initiation or interrupting propagation step. Secondary antioxidants quench singlet oxygen, decompose peroxides in non-radical species, chelate prooxidative metal ions, inhibit oxidative enzymes. Based on antioxidants’ reactivity, four lines of defense have been described: Preventative antioxidants, radical scavengers, repair antioxidants, and antioxidants relying on adaptation mechanisms. Carbon-based electrodes are largely employed in electroanalysis given their special features, that encompass large surface area, high electroconductivity, chemical stability, nanostructuring possibilities, facility of manufacturing at low cost, and easiness of surface modification. Largely employed methods encompass voltammetry, amperometry, biamperometry and potentiometry. Determination of key endogenous and exogenous individual antioxidants, as well as of antioxidant activity and its main contributors relied on unmodified or modified carbon electrodes, whose analytical parameters are detailed. Recent advances based on modifications with carbon-nanotubes or the use of hybrid nanocomposite materials are described. Large effective surface area, increased mass transport, electrocatalytical effects, improved sensitivity, and low detection limits in the nanomolar range were reported, with applications validated in complex media such as foodstuffs and biological samples.


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