Aptamer-based assays: strategies in the use of aptamers conjugated to magnetic micro- and nanobeads as recognition elements in food control

Mattarozzi, Monica; Toma, Lorenzo; Bertucci, Alessandro; Giannetto, Marco; Careri, Maria

doi:10.1007/s00216-021-03501-6

Cited by 11 publications

(15 citation statements)

References 45 publications

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“…Oligonucleotides that exhibit strong interactions with their target, and thus have high binding affinities, are characterized by a low K D . Generally, the reported K D values are in the picomolar to nanomolar range for proteins, and weaker-affinity interactions with K D in the nanomolar to micromolar range have been reported for small molecules [ 63 , 64 ]. Other quantitative metrics associated with the binding activity are selectivity and specificity [ 25 ].…”

Section: Aptamer-target Interactions—affinity Thermodynamic Parameter...mentioning

confidence: 99%

“…Beside affinity and selectivity, the determination of several other parameters such as buffer and ionic strength sensitivity should be performed when employing aptamers as bioreceptors, as it is known that experimental conditions such as buffer, ions, pH, temperature can influence the affinity of aptamers [ 64 , 66 , 67 ].…”

Section: Aptamer-target Interactions—affinity Thermodynamic Parameter...mentioning

confidence: 99%

“…The unfavorable contribution corresponds to entropic changes related to the decrease in the conformational entropy and the loss of the rotation and translation degrees of freedom of the aptamer-target complex in comparison with the free molecules [ 16 , 62 ]. Therefore, the intrinsic flexibility of an aptamer, which is very sensitive to its environment, causes an instability of its conformation, and thus a diminished affinity and selectivity for its specific target [ 8 , 64 ].…”

Section: Aptamer-target Interactions—affinity Thermodynamic Parameter...mentioning

confidence: 99%

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Critical Design Factors for Electrochemical Aptasensors Based on Target-Induced Conformational Changes: The Case of Small-Molecule Targets

et al. 2022

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Nucleic-acid aptamers consisting in single-stranded DNA oligonucleotides emerged as very promising biorecognition elements for electrochemical biosensors applied in various fields such as medicine, environmental, and food safety. Despite their outstanding features, such as high-binding affinity for a broad range of targets, high stability, low cost and ease of modification, numerous challenges had to be overcome from the aptamer selection process on the design of functioning biosensing devices. Moreover, in the case of small molecules such as metabolites, toxins, drugs, etc., obtaining efficient binding aptamer sequences proved a challenging task given their small molecular surface and limited interactions between their functional groups and aptamer sequences. Thus, establishing consistent evaluation standards for aptamer affinity is crucial for the success of these aptamers in biosensing applications. In this context, this article will give an overview on the thermodynamic and structural aspects of the aptamer-target interaction, its specificity and selectivity, and will also highlight the current methods employed for determining the aptamer-binding affinity and the structural characterization of the aptamer-target complex. The critical aspects regarding the generation of aptamer-modified electrodes suitable for electrochemical sensing, such as appropriate bioreceptor immobilization strategy and experimental conditions which facilitate a convenient anchoring and stability of the aptamer, are also discussed. The review also summarizes some effective small molecule aptasensing platforms from the recent literature.

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Section: Aptamer-target Interactions—affinity Thermodynamic Parameter...mentioning

confidence: 99%

Section: Aptamer-target Interactions—affinity Thermodynamic Parameter...mentioning

confidence: 99%

Section: Aptamer-target Interactions—affinity Thermodynamic Parameter...mentioning

confidence: 99%

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Critical Design Factors for Electrochemical Aptasensors Based on Target-Induced Conformational Changes: The Case of Small-Molecule Targets

et al. 2022

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“…Uncontrolled nonspecific interactions could influence the overall specificity and reproducibility of the methods, thus making the sample pretreatment a critical step for the analytical performances of these methods. Depending on the extraction procedure applied to food samples, the specific interaction between an aptamer and its target can be hindered by the presence of chemical or biological interferences that result in a reduction in the selectivity and sensitivity [ 56 ].…”

Section: Food Matrices and Sample Pretreatment For Portable Biosensorsmentioning

confidence: 99%

Luminescent Aptamer-Based Bioassays for Sensitive Detection of Food Allergens

et al. 2022

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The presence of hidden allergens in food products, often due to unintended contamination along the food supply chain (production, transformation, processing, and transport), has raised the urgent need for rapid and reliable analytical methods for detecting trace levels of such species in food products. Indeed, food allergens represent a high-risk factor for allergic subjects due to potentially life-threatening adverse reactions. Portable biosensors based on immunoassays have already been developed as rapid, sensitive, selective, and low-cost analytical platforms that can replace analyses with traditional bench-top instrumentation. Recently, aptamers have attracted great interest as alternative biorecognition molecules for bioassays, since they can bind a variety of targets with high specificity and selectivity, and they enable the development of assays exploiting a variety of transduction and detection technologies. In particular, aptasensors based on luminescence detection have been proposed, taking advantage of the development of ultrasensitive tracers and enhancers. This review aims to summarize and discuss recent efforts in the field of food allergen analysis using aptamer-based bioassays with luminescence detection.

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“…The most recent evolutions in analytical chemistry involve the combination of miniaturized devices to proper biological recognition elements designed to interact with high affinity and selectivity with the target compound. The biorecognition elements can exploit naturally occurring interactions, such as antibody-antigen, enzyme-substrate, and DNA-DNA binding [1][2][3], or they can be biomimetic synthetic receptors such as molecularly imprinted polymer or aptamers [4][5][6]. Aptamers are short ssDNA or ssRNA sequences synthetized from nucleic acids as biological units and selected in vitro through the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) iterative procedure, starting from an oligonucleotide library.…”

Section: Introductionmentioning

confidence: 99%

FFF-based high-throughput sequence shortlisting to support the development of aptamer-based analytical strategies

et al. 2022

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Aptamers are biomimetic receptors that are increasingly exploited for the development of optical and electrochemical aptasensors. They are selected in vitro by the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) procedure, but although they are promising recognition elements, for their reliable applicability for analytical purposes, one cannot ignore sample components that cause matrix effects. This particularly applies when different SELEX-selected aptamers and related truncated sequences are available for a certain target, and the choice of the aptamer should be driven by the specific downstream application. In this context, the present work aimed at investigating the potentialities of asymmetrical flow field-flow fractionation (AF4) with UV detection for the development of a screening method of a large number of anti-lysozyme aptamers towards lysozyme, including randomized sequences and an interfering agent (serum albumin). The possibility to work in native conditions and selectively monitor the evolution of untagged aptamer signal as a result of aptamer-protein binding makes the devised method effective as a strategy for shortlisting the most promising aptamers both in terms of affinity and in terms of selectivity, to support subsequent development of aptamer-based analytical devices. Graphical abstract

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Aptamer-based assays: strategies in the use of aptamers conjugated to magnetic micro- and nanobeads as recognition elements in food control

Cited by 11 publications

References 45 publications

Critical Design Factors for Electrochemical Aptasensors Based on Target-Induced Conformational Changes: The Case of Small-Molecule Targets

Critical Design Factors for Electrochemical Aptasensors Based on Target-Induced Conformational Changes: The Case of Small-Molecule Targets

Luminescent Aptamer-Based Bioassays for Sensitive Detection of Food Allergens

FFF-based high-throughput sequence shortlisting to support the development of aptamer-based analytical strategies

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