Sensitive gluten determination in gluten-free foods by an electrochemical aptamer-based assay

Amaya-González, Sonia; de‐los‐Santos‐Álvarez, Noemí; Miranda‐Ordieres, Arturo J.; Lobo-Castañón, Marı́a Jesús

doi:10.1007/s00216-015-8771-6

Cited by 44 publications

(30 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immobilization of the aptamer was found to be more deleterious in terms of affinity than the immobilization of the 33-mer peptide [17], so the latter was anchored through streptavidin-biotin interaction to the electrode surface. Among the aptamers raised against 33-mer [16], Gli 4 was selected because of its highest affinity and good performance in competitive magnetoassays [18]. Likewise, biotin was used as a tag for subsequent enzymatic labelling.…”

Section: Resultsmentioning

confidence: 99%

Disposable electrochemical aptasensor for gluten determination in food

López-López

Miranda‐Castro

de‐los‐Santos‐Álvarez

et al. 2017

Sensors and Actuators B: Chemical

Self Cite

View full text Add to dashboard Cite

Reliable detection of decreasing amounts of gluten in food is the only way of ensuring the safety of all coeliac patients. Results obtained with the method of choice, immunochemical assays, are not entirely comparable and many of them are sandwich assays that cannot recognize hydrolyzed proteins. In this work, we propose a competitive electrochemical sensor based on a recently described aptamer targeting the gliadin immunodominant peptide 33-mer that triggers the coeliac disease. The sensing layer is built on the surface of a screen-printed carbon electrode (SPCE) by adsorption of streptavidin and subsequent peptide immobilization. A competition between the peptide and gluten proteins from samples for a defined concentration of biotinylated Gli 4 aptamer is established. The aptamer bound to the peptide on the surface is finally measured after enzyme labelling and chronoamperometric detection of an enzymatically obtained electrochemically active product. This method is able to detect as low as 0.113 µg L-1 of gliadin, which corresponds to 380 µg kg-1 of gluten in food, taking all dilutions and conversion factors into consideration, with a reproducibility lower than 11%. The aptasensor was applied to food samples with gluten contents above and below the legislated threshold for gluten-free labelling in the EU, obtaining good agreement with the official R5 immunochemical method. mg kg -1Gluten Threshold gluten-free S P eaptasensor Graphical Abstract (for review) Highlights An electrochemical aptamer-based sensor on peptide-modified carbon screen-printed electrodes for gluten determination  Sensitivity is improved compared with reference enzyme-linked immunosorbent assays  The sensor was successfully applied to detect gluten in various food samples AbstractReliable detection of decreasing amounts of gluten in food is the only way of ensuring the safety of all coeliac patients. Results obtained with the method of choice, immunochemical assays, are not entirely comparable and many of them are sandwich assays that cannot recognize hydrolyzed proteins. In this work, we propose a competitive electrochemical sensor based on a recently described aptamer targeting the gliadin immunodominant peptide 33-mer that triggers the coeliac disease. The sensing layer is built on the surface of a screen-printed carbon electrode (SPCE) by adsorption of streptavidin and subsequent peptide immobilization. A competition between the peptide and gluten proteins from samples for a defined concentration of biotinylated Gli 4 aptamer is established. The aptamer bound to the peptide on the surface is finally measured after enzyme labelling and chronoamperometric detection of an enzymatically obtained electrochemically active product. This method is able to detect as low as 0.113 µg L -1 of gliadin, which corresponds to 380 µg kg -1 of gluten in food, taking all dilutions and conversion factors into consideration, with a reproducibility lower than 11%. The aptasensor was applied to food samples with gluten contents above and below the legislat...

show abstract

Section: Resultsmentioning

confidence: 99%

Disposable electrochemical aptasensor for gluten determination in food

López-López

Miranda‐Castro

de‐los‐Santos‐Álvarez

et al. 2017

Sensors and Actuators B: Chemical

Self Cite

View full text Add to dashboard Cite

show abstract

“…In the same context, Tran et al have successfully designed specific aptamers strands against food allergens including lysozyme [56] and Ara h 1 [59]. Several aptamer sequences for gluten were selected by two research groups, by taking as analytical target either the intact gliadin protein [60] or the immunodominant toxic 33-mer peptide [61]. Despite of the progress in the field of aptasensing, very few reports are available in literature for aptasensing detection of food allergens.…”

Section: Design Of Sensitive Biosensors For Allergen Based On Scrementioning

confidence: 99%

“…This method, however, has the disadvantage of low stability caused by desorption from the surface. The commonly used immobilization techniques to obtain a stable and active aptamer-functionalized surface include covalent coupling [39,42] click chemistry [43] and avidin-biotin affinity [61]. Chemisorption of the thiolated aptamer onto a gold electrode surface [52] could be envisaged as well, similarly to aptasensors for other analytes relevant for food safety.…”

Section: Design Of Sensitive Biosensors For Allergen Based On Scrementioning

confidence: 99%

“…Overall, magnetic beads proved to be versatile tools for electrochemical biosensing [82,87,88]. Association of screen-printed electrodes for cost-effective, easy detection with magnetic beads functionalized with high affinity, specific antibodies or aptamers delivered an advantageous approach for the detection of various analytes [88], including food allergens such as ovalbumin [82], β-lactoglobulin [89] Ara h 1 and Ara h 2 [90] and gluten [61]. An illustrative example of the potential and advantages of magnetoimmunosensors was provided by Ruiz-Montiel et al [90] for the simultaneous detection of Ara h 1 and Ara h 2 using dual screen-printed electrodes.…”

Section: Design Of Sensitive Biosensors For Allergen Based On Scrementioning

confidence: 99%

“…As per these strategies, HRP catalyses reactions such as (i) conversion of hydroquinone, in the presence of H 2 O 2 to benzoquinone (BQ) or (ii) oxidation of ,3,3’,5,5’-tetramethyl benzidine (TMB) in the presence of H 2 O 2 . Reduction of BQ [35,96] and detection of enzymatically oxidised TMB [61,66,97] by amperometry were linked to the amount of allergen present in the sample. …”

Section: Design Of Sensitive Biosensors For Allergen Based On Scrementioning

confidence: 99%

See 2 more Smart Citations

Electrochemical Affinity Biosensors Based on Disposable Screen-Printed Electrodes for Detection of Food Allergens

Vasilescu

Nunes

Hayat

et al. 2016

Sensors

View full text Add to dashboard Cite

Food allergens are proteins from nuts and tree nuts, fish, shellfish, wheat, soy, eggs or milk which trigger severe adverse reactions in the human body, involving IgE-type antibodies. Sensitive detection of allergens in a large variety of food matrices has become increasingly important considering the emergence of functional foods and new food manufacturing technologies. For example, proteins such as casein from milk or lysozyme and ovalbumin from eggs are sometimes used as fining agents in the wine industry. Nonetheless, allergen detection in processed foods is a challenging endeavor, as allergen proteins are degraded during food processing steps involving heating or fermentation. Detection of food allergens was primarily achieved via Enzyme-Linked Immuno Assay (ELISA) or by chromatographic methods. With the advent of biosensors, electrochemical affinity-based biosensors such as those incorporating antibodies and aptamers as biorecognition elements were also reported in the literature. In this review paper, we highlight the success achieved in the design of electrochemical affinity biosensors based on disposable screen-printed electrodes towards detection of protein allergens. We will discuss the analytical figures of merit for various disposable screen-printed affinity sensors in relation to methodologies employed for immobilization of bioreceptors on transducer surface.

show abstract