Immunochemical methods for detection of organophosphorus compounds

2020 ◽  
pp. 219-230
Author(s):  
Sergey Eremin

Organophosphorus compounds (OP) are found in environmental objects and food products. Due to their high toxicity and inhibition of cholinesterase activity, it is necessary to control residual amounts of OP. The most common methods for determining OP are gas and liquid chromatography with various detection methods. However, chromatographic analysis is lengthy, requires complex sample preparation and expensive equipment, which limits its use for screening a large number of samples and continuous monitoring of the content of OP. To detect the OP, it is necessary to use High Throughput Screening methods, using simple, fast and inexpensive analysis methods. Currently, immunochemical methods are increasingly used to determine OP. These methods are based on the recognition of the analyte (antigen) by specific receptors (antibodies) with the formation of the antigen-antibody complex and the measurement of the analytical signal generated by the immunochemical test system in response to complex formation, which leads to high sensitivity and specificity of the analysis.

Micromachines ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 390
Author(s):  
Maryia Drobysh ◽  
Almira Ramanaviciene ◽  
Roman Viter ◽  
Arunas Ramanavicius

The coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was proclaimed a global pandemic in March 2020. Reducing the dissemination rate, in particular by tracking the infected people and their contacts, is the main instrument against infection spreading. Therefore, the creation and implementation of fast, reliable and responsive methods suitable for the diagnosis of COVID-19 are required. These needs can be fulfilled using affinity sensors, which differ in applied detection methods and markers that are generating analytical signals. Recently, nucleic acid hybridization, antigen-antibody interaction, and change of reactive oxygen species (ROS) level are mostly used for the generation of analytical signals, which can be accurately measured by electrochemical, optical, surface plasmon resonance, field-effect transistors, and some other methods and transducers. Electrochemical biosensors are the most consistent with the general trend towards, acceleration, and simplification of the bioanalytical process. These biosensors mostly are based on the determination of antigen-antibody interaction and are robust, sensitive, accurate, and sometimes enable label-free detection of an analyte. Along with the specification of biosensors, we also provide a brief overview of generally used testing techniques, and the description of the structure, life cycle and immune host response to SARS-CoV-2, and some deeper details of analytical signal detection principles.


2022 ◽  
Vol 23 (2) ◽  
pp. 666
Author(s):  
Maryia Drobysh ◽  
Almira Ramanaviciene ◽  
Roman Viter ◽  
Chien-Fu Chen ◽  
Urte Samukaite-Bubniene ◽  
...  

Monitoring and tracking infection is required in order to reduce the spread of the coronavirus disease 2019 (COVID-19), induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To achieve this goal, the development and deployment of quick, accurate, and sensitive diagnostic methods are necessary. The determination of the SARS-CoV-2 virus is performed by biosensing devices, which vary according to detection methods and the biomarkers which are inducing/providing an analytical signal. RNA hybridisation, antigen-antibody affinity interaction, and a variety of other biological reactions are commonly used to generate analytical signals that can be precisely detected using electrochemical, electrochemiluminescence, optical, and other methodologies and transducers. Electrochemical biosensors, in particular, correspond to the current trend of bioanalytical process acceleration and simplification. Immunosensors are based on the determination of antigen-antibody interaction, which on some occasions can be determined in a label-free mode with sufficient sensitivity.


2007 ◽  
Vol 14 (5) ◽  
pp. 505-509 ◽  
Author(s):  
Erik Avaniss-Aghajani ◽  
Sophia Berzon ◽  
Arlen Sarkissian

ABSTRACT The advent of multiplexed bead assays in recent years has introduced a new dimension of testing for complex diseases such as lupus, which can involve multiple autoantibodies. The ability to rapidly identify multiple autoantibodies, with high sensitivity and specificity in an automated fashion, is highly attractive. The aim of this study was to assess the performance and clinical value of multiplexed bead-based (AtheNA Multi-Lyte ANA-II test system) immunoassays both by comparing the results with those achieved by indirect fluorescent-antibody assay (IFA) or conventional enzyme immunoassays (EIAs) and by independent identification of autoantibodies in well-characterized samples. To achieve this goal, 984 samples were tested for seven analytes (SS/A, SS/B, Sm, RNP, Scl-70, double-stranded DNA [dsDNA], and centromere B) in both traditional and bead-based assays. The average concordance for the different analytes was 91%, ranging from 81% (dsDNA) to 97% (centromere B). The average relative specificity and sensitivity for the analytes were also high, 92% and 81%, respectively. An examination of 93 “normal controls” demonstrated a 7% false-positive rate, which was comparable to IFA. Percentages of different autoantibodies found in patients with a variety of disease conditions (34 with calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia; 41 with mixed connective tissue disease; 24 with scleroderma; and 35 with Sjogren's syndrome) were well within the range expected from each group. A scrutiny of results from AtheNA and EIA and Farr results for 185 systemic lupus erythematosus samples revealed comparable results by both methods, with the exception of SS/A and dsDNA, where AtheNA had a higher percentage of SS/A-positive results compared to EIA (51% versus 29%) and a lower percentage of dsDNA-positive results (18% versus 28% at a cutoff of 5 IU/ml).


2013 ◽  
Vol 690-693 ◽  
pp. 1449-1454
Author(s):  
Yuan Wang ◽  
Hui Juan Yu ◽  
Bei Lei Qian ◽  
You Qiong Cai ◽  
Dong Mei Huang ◽  
...  

Gold immunochromatography assay (GICA) technique has the following characteristics: rapid and simple, high sensitivity, good specificity, no auxiliary equipment, ease of interpreting results, and satisfactory stability. The technique has become one of the most rapid and sensitive immunological detection methods, which is widely used in medical, biological and other fields. The article focuses on the basic principles and technical characteristics of GICA, and briefly describes the applications and future perspectives in the rapid detection of aquatic products.


Author(s):  
Dong-Jiunn Jeffery Truong ◽  
Teeradon Phlairaharn ◽  
Bianca Eßwein ◽  
Christoph Gruber ◽  
Deniz Tümen ◽  
...  

AbstractExpression of exon-specific isoforms from alternatively spliced mRNA is a fundamental mechanism that substantially expands the proteome of a cell. However, conventional methods to assess alternative splicing are either consumptive and work-intensive or do not quantify isoform expression longitudinally at the protein level. Here, we therefore developed an exon-specific isoform expression reporter system (EXSISERS), which non-invasively reports the translation of exon-containing isoforms of endogenous genes by scarlessly excising reporter proteins from the nascent polypeptide chain through highly efficient, intein-mediated protein splicing. We applied EXSISERS to quantify the inclusion of the disease-associated exon 10 in microtubule-associated protein tau (MAPT) in patient-derived induced pluripotent stem cells and screened Cas13-based RNA-targeting effectors for isoform specificity. We also coupled cell survival to the inclusion of exon 18b of FOXP1, which is involved in maintaining pluripotency of embryonic stem cells, and confirmed that MBNL1 is a dominant factor for exon 18b exclusion. EXSISERS enables non-disruptive and multimodal monitoring of exon-specific isoform expression with high sensitivity and cellular resolution, and empowers high-throughput screening of exon-specific therapeutic interventions.


2021 ◽  
Vol 368 (6) ◽  
Author(s):  
Liwen Zhang ◽  
Qingyu Lv ◽  
Yuling Zheng ◽  
Xuan Chen ◽  
Decong Kong ◽  
...  

ABSTRACT T-2 is a common mycotoxin contaminating cereal crops. Chronic consumption of food contaminated with T-2 toxin can lead to death, so simple and accurate detection methods in food and feed are necessary. In this paper, we establish a highly sensitive and accurate method for detecting T-2 toxin using AlphaLISA. The system consists of acceptor beads labeled with T-2-bovine serum albumin (BSA), streptavidin-labeled donor beads and biotinylated T-2 antibodies. T-2 in the sample matrix competes with T-2-BSA for antibodies. Adding biotinylated antibodies to the test well followed by T-2 and T-2-BSA acceptor beads yielded a detection range of 0.03–500 ng/mL. The half-maximal inhibitory concentration was 2.28 ng/mL and the coefficient of variation was <10%. In addition, this method had no cross-reaction with other related mycotoxins. This optimized method for extracting T-2 from food and feed samples achieved a recovery rate of approximately 90% in T-2 concentrations as low as 1 ng/mL, better than the performance of a commercial ELISA kit. This competitive AlphaLISA method offers high sensitivity, good specificity, good repeatability and simple operation for detecting T-2 toxin in food and feed.


Pharmaceutics ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1019
Author(s):  
Teresa von Linde ◽  
Gzona Bajraktari-Sylejmani ◽  
Walter E. Haefeli ◽  
Jürgen Burhenne ◽  
Johanna Weiss ◽  
...  

The peptide transporter PEPT-1 (SLC15A1) plays a major role in nutritional supply with amino acids by mediating the intestinal influx of dipeptides and tripeptides generated during food digestion. Its role in the uptake of small bioactive peptides and various therapeutics makes it an important target for the investigation of the systemic absorption of small peptide-like active compounds and prodrug strategies of poorly absorbed therapeutics. The dipeptide glycyl-sarcosine (Gly-Sar), which comprises an N-methylated peptide bond that increases stability against enzymatic degradation, is widely utilized for studying PEPT-1-mediated transport. To support experiments on PEPT-1 inhibitor screening to identify potential substrates, we developed a highly sensitive Gly-Sar quantification assay for Caco-2 cell lysates with a dynamic range of 0.1 to 1000 ng/mL (lower limit of quantification 0.68 nM) in 50 µL of cell lysate. The assay was validated following the applicable recommendations for bioanalytic method validation of the FDA and EMA. Sample preparation and quantification were established in 96-well cell culture plates that were also used for the cellular uptake studies, resulting in a rapid and robust screening assay for PEPT-1 inhibitors. This sample preparation principle, combined with the high sensitivity of the UPLC-MS/MS quantification, is suitable for screening assays for PEPT-1 inhibitors and substrates in high-throughput formats and holds the potential for automation. Applicability was demonstrated by IC50 determinations of the known PEPT-1 inhibitor losartan, the known substrates glycyl-proline (Gly-Pro), and valaciclovir, the prodrug of aciclovir, which itself is no substrate of PEPT-1 and consequently showed no inhibition in our assay.


2008 ◽  
Vol 1133 ◽  
Author(s):  
Brian T. Cunningham ◽  
Leo Chan ◽  
Patrick C. Mathias ◽  
Nikhil Ganesh ◽  
Sherine George ◽  
...  

Abstract Photonic crystal surfaces represent a class of resonant optical structures that are capable of supporting high intensity electromagnetic standing waves with near-field and far-field properties that can be exploited for high sensitivity detection of biomolecules and cells. While modulation of the resonant wavelength of a photonic crystal by the dielectric permittivity of adsorbed biomaterials enables label-free detection, the resonance can also be tuned to coincide with the excitation wavelength of common fluorescent tags - including organic molecules and semiconductor quantum dots. Photonic crystals are also capable of efficiently channeling fluorescent emission into a preferred direction for enhanced extraction efficiency. Photonic crystals can be designed to support multiple resonant modes that can perform label free detection, enhanced fluorescence excitation, and enhanced fluorescence extraction simultaneously on the same device. Because photonic crystal surfaces may be inexpensively produced over large surface areas by nanoreplica molding processes, they can be incorporated into disposable labware for applications such as pharmaceutical high throughput screening. In this talk, the optical properties of surface photonic crystals will be reviewed and several applications will be described, including results from screening a 200,000-member chemical compound library for inhibitors of protein-DNA interactions, gene expression microarrays, and high sensitivity of protein biomarkers.


2012 ◽  
Vol 532-533 ◽  
pp. 398-402
Author(s):  
Yu Lan Wei ◽  
Bing Li ◽  
Sui Ying Jin ◽  
Kai Kai Chen

An integrated system to measure mechanical functions of material or structure is introduced. This system is able to provide more detection methods and experiment environments. And it can discover the characteristics and mechanisms of damnification and breakage in materials, considering the effects of loading and environments. Material functions were analyzed in many aspects, including loading, strain, light, sound, temperature and infrared to ensure the safety of materials and configuration of unmanned plane. Current study has laid a foundation for realizing optimal design of unmanned plane. In this paper, theory, components, and function of the system were discussed, as well as signal acquisition and analysis.


2012 ◽  
Vol 229-231 ◽  
pp. 1423-1426
Author(s):  
Yong Zheng Wang ◽  
Lei Jiang ◽  
Mao Zhen Yue ◽  
Su Fang Bian

In this paper, three detection methods were analyzed to determine the chlorine precipitation from the flue gas: Silver nitrate volumetric analysis (SNVA), Volhard method (VM) and Mercuric thiocyanate spectrophotometric method (MTSM). Results indicated that SNVA and VM were simple and convenient, but unsuitable for the detection of blend fuel due to the high lower limit. MTSM was suitable for biomass and coal co-combustion for its low lower limit and high sensitivity. In addition, MTSM needed less sample solution, which made it more suitable for multiple measurements to reduce errors, and the maximum proportional error was only 3.5%.


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