Immobilization of Aptamers on Substrates

Girolamo, Annalisa De; McKeague, Maureen; Pascale, Michelangelo; Cortese, Marina; DeRosa, Maria C.

doi:10.1002/9783527806799.ch3

Cited by 15 publications

(29 citation statements)

References 114 publications

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“…These technologies are costly and not always available in standard molecular biology laboratories where SELEX is typically performed [31]. Furthermore, these challenges are particularly problematic in the context of small molecule-binding aptamers, like mycotoxins, because most affinity binding assays are not sufficiently sensitive to measure the interaction of low molecular weight targets with the aptamer [33,34]. To overcome these limitations, several computational methods have been described in literature with the aim of streamlining the screening and selection of the aptamer during or post-SELEX [35,36].…”

Section: Introductionmentioning

confidence: 99%

An In-Silico Pipeline for Rapid Screening of DNA Aptamers against Mycotoxins: The Case-Study of Fumonisin B1, Aflatoxin B1 and Ochratoxin A

et al. 2020

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Aptamers are single-stranded oligonucleotides selected by SELEX (Systematic Evolution of Ligands by EXponential Enrichment) able to discriminate target molecules with high affinity and specificity, even in the case of very closely related structures. Aptamers have been produced for several targets including small molecules like mycotoxins; however, the high affinity for their respective target molecules is a critical requirement. In the last decade, the screening through computational methods of aptamers for their affinity against specific targets has greatly increased and is becoming a commonly used procedure due to its convenience and low costs. This paper describes an in-silico approach for rapid screening of ten ssDNA aptamer sequences against fumonisin B1 (FB1, n = 3), aflatoxin B1 (AFB1, n = 2) and ochratoxin A (OTA, n = 5). Theoretical results were compared with those obtained by testing the same aptamers by fluorescent microscale thermophoresis and by magnetic beads assay for their binding affinity (KD) revealing a good agreement.

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Section: Introductionmentioning

confidence: 99%

An In-Silico Pipeline for Rapid Screening of DNA Aptamers against Mycotoxins: The Case-Study of Fumonisin B1, Aflatoxin B1 and Ochratoxin A

et al. 2020

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“…Oligonucleotide aptamers can be easily modified by convenient functional groups like amino, azido, thiol, or biotin, which can be incorporated at the 3′ and/or 5′ ends during synthesis, in order to facilitate their labeling or immobilization onto different platforms. Several studies indicated that the 3′-end is more suitable for surface immobilization, as it may confer resistance to exonucleases [ 19 , 90 ]. Nevertheless, the 5′-end is frequently used for attachment, considering that modifications at this position are performed at the final stage of oligonucleotide synthesis, which improves yield and simplifies purification [ 19 ].…”

Section: Strategies In Electrochemical Aptasensor Design: Aptamer Immobilization and Electrochemical Signal Generationmentioning

confidence: 99%

“…Several studies indicated that the 3′-end is more suitable for surface immobilization, as it may confer resistance to exonucleases [ 19 , 90 ]. Nevertheless, the 5′-end is frequently used for attachment, considering that modifications at this position are performed at the final stage of oligonucleotide synthesis, which improves yield and simplifies purification [ 19 ]. The additional incorporation of spacer groups between the oligonucleotide sequence and the attachment point can improve the aptamers’ conformational flexibility, ultimately leading to an enhanced sensitivity [ 19 , 76 , 91 , 92 ].…”

Section: Strategies In Electrochemical Aptasensor Design: Aptamer Immobilization and Electrochemical Signal Generationmentioning

confidence: 99%

“…Nucleic acid aptamers are single-stranded DNA (ssDNA) or RNA sequences isolated from a vast library of synthetic random oligonucleotides via an automated process known as Systematic Evolution of Ligands by Exponential Enrichment (SELEX) [ 16 ]. Aptamer molecules can compete with antibodies in terms of affinity and specificity, have superior chemical stability, and, unlike antibodies, can be reversibly denatured in order to release the target analyte and enable device reusability [ 17 , 18 , 19 ]. As aptamers can be obtained through custom synthesis at reasonable prices, they are increasingly replacing antibodies in the fabrication of affinity biosensors [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

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The Role of Aryldiazonium Chemistry in Designing Electrochemical Aptasensors for the Detection of Food Contaminants

Raicopol

Pilan

2021

Materials

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Food safety monitoring assays based on synthetic recognition structures such as aptamers are receiving considerable attention due to their remarkable advantages in terms of their ability to bind to a wide range of target analytes, strong binding affinity, facile manufacturing, and cost-effectiveness. Although aptasensors for food monitoring are still in the development stage, the use of an electrochemical detection route, combined with the wide range of materials available as transducers and the proper immobilization strategy of the aptamer at the transducer surface, can lead to powerful analytical tools. In such a context, employing aryldiazonium salts for the surface derivatization of transducer electrodes serves as a simple, versatile and robust strategy to fine-tune the interface properties and to facilitate the convenient anchoring and stability of the aptamer. By summarizing the most important results disclosed in the last years, this article provides a comprehensive review that emphasizes the contribution of aryldiazonium chemistry in developing electrochemical aptasensors for food safety monitoring.

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“…Nanomaterial-based microcantilever (MC) biosensors have good potential in advanced biosensor technology as they offer a high surface-to-volume ratio [27][28][29], so they are able to mediate faster and with a higher kinetic electron transfer [30,31]. The size of nanomaterials can also facilitate effective interaction with targeted biomolecules.…”

Section: Introductionmentioning

confidence: 99%

Synthesis, Characterisation and Cytotoxicity of Gold Nanowires for Ultra-Sensitive Biosensor Development

Lah

Gray

Trigueros³

2020

Preprint

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With the long-term goal of developing an ultra-sensitive microcantilever-based nanobiosensor for versatile biomarker detection, new controlled bioreceptor-analytes systems are being explored to overcome the disadvantages of conventional ones. Gold nanowires (AuNWs) have been used as a probe to overcome the tolerance problem that occurs in response to changes in environmental conditions. However, the cytotoxicity of AuNWs is still unclear. Here, we examined the cytotoxicity of AuNWs systems using both commercial and as-synthesised AuNWs. In vitro experiments show that commercial AuNWs with an average quoted length of 6 µm are highly toxic against Gram-negative Escherichia coli (E. coli) at 50 µg/ml. However, this toxicity is due to the presence of CTAB surfactant no by the nanostructure. Conversely, the as-synthesised AuNWs with an average length of 9.5 µm show non-cytotoxicity even at the maximum viable concentration (330 µg/ml). These findings may lead to the development of potentially life-saving cytotoxicity -free nanobiosensors for an early diagnostic of potential diseases.

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Immobilization of Aptamers on Substrates

Cited by 15 publications

References 114 publications

An In-Silico Pipeline for Rapid Screening of DNA Aptamers against Mycotoxins: The Case-Study of Fumonisin B1, Aflatoxin B1 and Ochratoxin A

An In-Silico Pipeline for Rapid Screening of DNA Aptamers against Mycotoxins: The Case-Study of Fumonisin B1, Aflatoxin B1 and Ochratoxin A

The Role of Aryldiazonium Chemistry in Designing Electrochemical Aptasensors for the Detection of Food Contaminants

Synthesis, Characterisation and Cytotoxicity of Gold Nanowires for Ultra-Sensitive Biosensor Development

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