Selection, characterization and application of aptamers targeted to Aflatoxin B2

Ma, Xiaoyuan; Wang, Wenfeng; Chen, Xiujuan; Xia, Yu; Duan, Nuo; Wu, Shijia; Wang, Zhouping

doi:10.1016/j.foodcont.2014.07.037

Cited by 46 publications

(15 citation statements)

References 32 publications

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“…Therefore, immobilization of targets and their analogs onto the matrices by complex chemical reactions might alter the native confirmations and possibly lead to reduction in aptamer affinity (Le et al, 2010; McKeague et al, 2010, 2014; Chen et al, 2013, 2014; Ma et al, 2014, 2015; Malhotra et al, 2014). In present report, we describe a modified immunoaffinity based SELEX procedure for selection of aptamers against AFB1.…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, aptamers are capable of retaining their reactivity under a wide range of environmental conditions (Famulok et al, 2007; Iliuk et al, 2011). Recently, various high throughput SELEX technologies have been investigated for development of aptamers against mycotoxins such as Aflatoxins (AFB1, AFB2, and AFM1), Fumonisin B1 (FB1), Zearalenone (ZEA), T2 toxin and Ochratoxin A (OTA; Le et al, 2010; McKeague et al, 2010, 2014; Chen et al, 2013, 2014; Ma et al, 2014, 2015; Malhotra et al, 2014). In most of these reports, mycotoxins are coupled to magnetic nanoparticles, sepharose, agarose based resin, or streptavidin beads prior to selection and screening of aptamers.…”

Section: Introductionmentioning

confidence: 99%

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Immuno Affinity SELEX for Simple, Rapid, and Cost-Effective Aptamer Enrichment and Identification against Aflatoxin B1

et al. 2016

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Aflatoxins are naturally occurring mycotoxins that contaminate food and agro commodities, leading to acute and chronic health conditions in human and animals. In the present work, an attempt was made to generate high-affinity single stranded DNA aptamers that specifically bind to Aflatoxin B1 (AFB1) by a modified Systemic Evolution of Ligands by Exponential Enrichment (SELEX) procedure with the aid of Immunoaffinity columns. Ten rounds of SELEX and alternating three counter SELEX rounds with a cocktail of related and other mycotoxins were performed to enhance the specificity. Resultant 105 aptamers were clustered into 12 groups according to their primary sequence homology. Candidates with lowest Gibbs free energy (dG value) and unique stem loop structures were selected for further characterization. Aptamers, AFLA5, AFLA53, and AFLA71 exhibiting lower Kd values (50.45 ± 11.06, 48.29 ± 9.45, and 85.02 ± 25.74 nM) were chosen for development of ELONA and determination of purification ability of toxin. The detection limit (LOD) of AFLA5 and AFLA71 was 20 and 40 ng/ml, respectively. HPLC analysis implied that selected aptamers were able to recover and quantify 82.2 to 96.21% (LOQ – 53.74 ng) and 78.3 to 94.22% (LOQ – 66.75 ng) of AFB1 from spiked corn samples, respectively. These findings indicate, immunoaffinity based SELEX can pave an alternative approach to screen aptamers against mycotoxin detection and purification.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Immuno Affinity SELEX for Simple, Rapid, and Cost-Effective Aptamer Enrichment and Identification against Aflatoxin B1

et al. 2016

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“…AFB1 aptamer was first selected and patented by Neoventures Biotechnology Inc. (London, ON, Canada) [90]. Then, Wang’s group identified specific aptamers for AFB1 and B2 [91], while AFM1 aptamer has been selected by Malhotra et al [92]. After selection, these aptamers have been used, in many reports, as biorecognition elements in optical and electrochemical label-free aptasensing strategies.…”

Section: Label-free Aptasensors For Myctotoxin Determinationmentioning

confidence: 99%

Label-Free Aptasensors for the Detection of Mycotoxins

Rhouati

Catanante

Nunes

et al. 2016

Sensors

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Various methodologies have been reported in the literature for the qualitative and quantitative monitoring of mycotoxins in food and feed samples. Based on their enhanced specificity, selectivity and versatility, bio-affinity assays have inspired many researchers to develop sensors by exploring bio-recognition phenomena. However, a significant problem in the fabrication of these devices is that most of the biomolecules do not generate an easily measurable signal upon binding to the target analytes, and signal-generating labels are required to perform the measurements. In this context, aptamers have been emerged as a potential and attractive bio-recognition element to design label-free aptasensors for various target analytes. Contrary to other bioreceptor-based approaches, the aptamer-based assays rely on antigen binding-induced conformational changes or oligomerization states rather than binding-assisted changes in adsorbed mass or charge. This review will focus on current designs in label-free conformational switchable design strategies, with a particular focus on applications in the detection of mycotoxins.

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“…Recently, aptamer sequences possessing a high affinity for AFs (different structural analogs) have been reported in either publications or patents (Table 3) [66][67][68][69][70]. In the present scenario, the problems associated with the analysis of complex samples, such as blood, serum, and cellular extracts, have been solved using electrochemical biosensors [71,72].…”

Section: Aptamer-based Sensing Strategies For Aflatoxins (Afs) Detectionmentioning

confidence: 99%

Recent Advances in Electrochemical-Based Sensing Platforms for Aflatoxins Detection

et al. 2016

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Abstract:Mycotoxin are small (MW~700 Da), toxic secondary metabolites produced by fungal species that readily colonize crops and contaminate them at both pre-and post-harvesting. Among all, aflatoxins (AFs) are mycotoxins of major significance due to their presence in common food commodities and the potential threat to human health worldwide. Based on the severity of illness and increased incidences of AFs poisoning, a broad range of conventional and analytical detection techniques that could be useful and practical have already been reported. However, due to the variety of structural analogous of these toxins, it is impossible to use one common technique for their analysis. Numerous recent research efforts have been directed to explore alternative detection technologies. Recently, immunosensors and aptasensors have gained promising potential in the area of sample preparation and detection systems. These sensors offer the advantages of disposability, portability, miniaturization, and on-site analysis. In a typical design of an aptasensor, an aptamer (ssDNA or RNA) is used as a bio-recognition element either integrated within or in intimate association with the transducer surface. This review paper is focused on the recent advances in electrochemical immunoand aptasensing platforms for detection of AFs in real samples.

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Selection, characterization and application of aptamers targeted to Aflatoxin B2

Cited by 46 publications

References 32 publications

Immuno Affinity SELEX for Simple, Rapid, and Cost-Effective Aptamer Enrichment and Identification against Aflatoxin B1

Immuno Affinity SELEX for Simple, Rapid, and Cost-Effective Aptamer Enrichment and Identification against Aflatoxin B1

Label-Free Aptasensors for the Detection of Mycotoxins

Recent Advances in Electrochemical-Based Sensing Platforms for Aflatoxins Detection

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