The Enzyme-Linked Immunosorbent Assay

Drijvers, Jefte M.; Awan, Imad M.; Perugino, Cory A.; Rosenberg, Ian M.; Pillai, Shiv

doi:10.1016/b978-0-12-803077-6.00007-2

Cited by 20 publications

(14 citation statements)

References 9 publications

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“…Enzyme-linked immunosorbent assay (ELISA): Immunoassays, the most popular type of which is ELISA, provide quantification and identification of various antigens [21]. ELISAs involve simple enzyme assays with specificity of antibodies using antigens or antibodies linked to an easily assayed enzyme.…”

Section: Chapter Outlinementioning

confidence: 99%

“…ELISAs involve simple enzyme assays with specificity of antibodies using antigens or antibodies linked to an easily assayed enzyme. The basic principle of ELISA technique is radioimmunoassay process that involves a tagged antibody in order to detect antigens [14,21]. An enzyme bound with a substrate, a fluorescent molecule, or a radioactive isotope can be described as the tag [14].…”

Section: Chapter Outlinementioning

confidence: 99%

“…On the other hand, ELISA tests can sometimes be quite expensive due to the cost of reagents being used [23]. Direct ELISA, indirect ELISA, sandwich ELISA, and competitive ELISA are the different types of ELISA tests which are used for different scientific fields [14,21,22].…”

Section: Chapter Outlinementioning

confidence: 99%

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Electrochemical virus detections with nanobiosensors

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Section: Chapter Outlinementioning

confidence: 99%

Section: Chapter Outlinementioning

confidence: 99%

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“…Immunoassays are widely used for screening and preliminary analysis of mycotoxins due to their advantages of high throughput, economy, and simplicity (Zhao et al., 2019). Based on the specific affinity between an antigen and an antibody (Zhang et al., 2022), major immunoassay methods include enzyme‐linked immunosorbent assay (ELISA) (Drijvers et al., 2017), lateral flow assay (LFAs) (Charlermroj et al., 2021), fluorescence polarization assay (Huang et al., 2020), and radioimmunoassay (Ayoub et al., 2016). Notably, LFAs for detecting mycotoxins have matured to commercialization due to their simplicity, portability, sensitivity, selectivity, and rapidity (Pan et al., 2020).…”

Section: Introductionmentioning

confidence: 99%

Improvement of the sensitivity of lateral flow systems for detecting mycotoxins: Up‐to‐date strategies and future perspectives

Li,

Jallow,

Nidiaye

et al. 2023

Comp Rev Food Sci Food Safe

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Mycotoxins are dangerous human and animal health‐threatening secondary fungal metabolites that can be found in various food and agricultural products. Several countries have established regulations to restrict their presence in food and agricultural products destined for human and animal consumption. Consequently, the need to develop highly sensitive and smart detection systems was recognized worldwide. Lateral flow assay possesses the advantages of easy operation, rapidity, stability, accuracy, and specificity, and it plays an important role in the detection of mycotoxins. Nevertheless, strategies to comprehensively improve the sensitivity of lateral flow assay to mycotoxins in food have rarely been highlighted and discussed. In this article, a comprehensive overview was presented on the application of lateral flow assay in mycotoxin detection in food samples by highlighting the principle of lateral flow assay, presenting a detailed discussion on various analytical performance‐improvement strategies, such as the development of high‐affinity recognition reagents, immunogen immobilization methods, and signal amplification. Additionally, a detailed discussion on the various signal analyzers and interpretation approaches was provided. Finally, current hurdles and future perspectives on the application of lateral flow assay in the detection of mycotoxins were discussed.

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“…Enzyme-linked immunosorbent assays (ELISA) adapted to different formats, e.g., direct, indirect, sandwich, and competitive ELISA are routine biochemical assays involving antigen–antibody binding for high-throughput and ultrasensitive detection of low and high molecular mass compounds in a variety of research fields including clinical, environmental, , and food analysis . In fact, recent trends geared toward the development of ELISA tests for emerging organic pollutants, e.g., pharmaceuticals, personal care products, and endocrine-disrupting chemicals, in environmental waters. , The standard ELISA protocol involves the colorimetric detection of the biochemical product of the prior enzymatic reaction, e.g., hydrogen peroxide, by resorting to organic chromophores, such as 3,3′,5,5′-tetramethylbenzidine (TMB), o -phenylenediamine (OPD), and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS). ,,− However, colorimetric competitive and sandwich ELISA sensing platforms may have limited sensitivity for determination of low-molecular mass pollutants at environmentally relevant levels because the detection is merely based on the color measurement of the resulting solution by conventional photometric analysis. Especially, this comes true for detecting pollutants in marine ecosystems that are found at low ng L –1 (ppt) levels.…”

mentioning

confidence: 99%

Reliable Sensing Platform for Plasmonic Enzyme-Linked Immunosorbent Assays Based on Automatic Flow-Based Methodology

et al. 2019

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Plasmonic enzyme-linked immunosorbent assays (ELISA) using the localized surface plasmon resonance (LSPR) of metal nanoparticles has emerged as an appealing alternative to conventional ELISA counterparts for ultrasensitive naked-eye detection of biomolecules and small contaminants. However, batchwise plasmonic ELISA involving end-point detection lacks ruggedness inasmuch as the generation or etching of NP is greatly dependent on every experimental parameter of the analytical workflow. To tackle the above shortcomings, this paper reports on an automatic flow methodology as a reliable detection scheme of hydrogen peroxide related enzymatic bioassays for ultrasensitive detection of small molecules. Here, a competitive ELISA is combined with the in-line generation of plasmonic gold nanoparticles (AuNPs) followed by the real-time monitoring of the NP nucleation and growth rates and size distribution using a USB miniaturized photometer. Glucose oxidase was labeled to the secondary antibody and yielded hydrogen peroxide that acted as the measurand and the reducing agent of the Au(III)/citrate system in the flow network. High-throughput plasmonic assays were feasible by assembling a hybrid flow system composed of two microsyringe pumps, a perfluoroalkoxy alkane reaction coil, and a 26-port multiposition valve and operated under computer-controllable flow conditions. The ultratrace determination of diclofenac in high matrix samples, e.g., seawater, without any prior sample treatment was selected as a proof-of-concept application of the flow-based platform for determination of emerging contaminants via plasmonic ELISA. The detection limit (0.001 μg L–1) was 1 order of magnitude lower than that endorsed by the first EU Watch List for diclofenac as a potentially emerging contaminant in seawater and also than that of a conventional colorimetric ELISA, which in turn is inappropriate for determination of diclofenac in seawater at the levels endorsed by the EU regulation. The proposed automatic fluidic approach is characterized by the reproducible timing in AuNPs nucleation and growth along with the unsupervised LSPR absorbance detection of AuNPs with a dynamic range for diclofenac spanning from 0.01 to 10 μg L–1. Repeatability and intermediate precision (given as normalized signal readouts) in seawater were <4% and <14%, respectively, as compared to RSDs as high as 30% as obtained with the batchwise plasmonic ELISA counterpart.

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The Enzyme-Linked Immunosorbent Assay

Cited by 20 publications

References 9 publications

Electrochemical virus detections with nanobiosensors

Electrochemical virus detections with nanobiosensors

Improvement of the sensitivity of lateral flow systems for detecting mycotoxins: Up‐to‐date strategies and future perspectives

Reliable Sensing Platform for Plasmonic Enzyme-Linked Immunosorbent Assays Based on Automatic Flow-Based Methodology

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