Detection of Allergenic Lysozyme during Winemaking with an Electrochemical Aptasensor

Abstract

In this work we investigate the performance of a simple, disposable electrochemical aptasensor for lysozyme and its usefulness for monitoring the allergen risk along wine production. The sensor relies on screen‐printed gold electrodes modified with gold nanoparticles as the electrochemical transducer, with detection by cyclic voltammetry. This simple method is characterized by a detection limit of 0.32 μg.mL−1 lysozyme and a linear range of 1–10 μg.mL−1, being appropriate for the analysis of lysozyme‐treated wines. Several white wines where sulphur dioxide was partially replaced by lysozyme were produced and analyzed with the aptasensor at critical stages during wine production. The results obtained with the aptasensor were moreover compared with those recorded in parallel by a standard method, high performance liquid chromatography (HPLC). The specific advantages brought by the use of nanomaterial and the limitations of the sensor are discussed. The sensor allowed evaluating the effect of various technological steps along wine production on the content of lysozyme.

Cutting-Edge Advances in Electrochemical Affinity Biosensing at Different Molecular Level of Emerging Food Allergens and Adulterants

“…The methods developed making use of aptamers for the determination of protein allergens such as gluten or lysozyme is also noteworthy. Table 2 shows as a label-free aptasensor has been developed for the detection of lysozyme using a direct format implemented on electrodes nanostructured with AuNPs [33]. The methods developed for the detection of gluten, requiring a high sensitivity, involved competitive formats between gluten proteins (gliadin) and a synthetic biotinylated peptide immobilized on the surface of a SPCE (Figure 6a,b) [16] or Strep-MBs [34].…”

“…Immunosensors are the most widely used biosensors to date in this field. However, the well-known antibodies limitations such as their in vivo production, low stability and possible cross-reactivity with non-target compounds of similar structure, has led to explore the use of synthetic receptors such as aptamers [32][33][34], peptides [34], nucleic acids and oligonucleotide-switching probes [35].…”

Progress in Electrochemical (Bio)Sensors for Monitoring Wine Production

“…With regard to the proteins used as processing aids in wine production, several electrochemical biosensors as well as bioassays coupled with electrochemical detection were proposed for the detection of milk and egg proteins (casein, B-lactoglobulin, ovalbumin, lysozyme) [107]. However, only a few of these sensors were tested so far with wines: e.g., optical biosensors for lysozyme [108][109][110] and ovalbumin [111] and electrochemical ones for lysozyme [10,110,112]. The development of biosensors for lysozyme was prompted by the availability of several aptamer sequences specific for lysozyme which can bind lysozyme with high affinity (affinity constants in the nM range).…”

“…Since the first report on a selected lysozyme aptamer in 2001, lysozyme was a preferred model analyte when developing novel biosensing strategies involving different types of detection formats (e.g., direct, competitive, sandwich). Among the various electrochemical biosensors [113], several were applied for wine analysis and used voltamperometric detection [10,112], as illustrated in Figure 11. Ocana et al developed a biosensor that combined a capture aptamer immobilized on the electrode surface with a detection antibody, in a sandwich detection format [112].…”

“…However, voltammetry performed at bare or modified electrodes remains a very powerful tool for analyzing wine composition [4]. In particular, applications of cyclic voltammetry include measuring the content of total polyphenols and total antioxidant capacity of wines [5][6][7], free sulphur dioxide [8,9] or the allergenic protein lysozyme [10]. In addition to CV, DPV [11,12], SWV [13], and LSV [14] have been shown to be very useful for obtaining qualitative and quantitative information regarding the poliphenolic composition [11,13] and the content of sulphites [14] or ascorbic acid [12] in wines.Voltammetric responses were correlated with organoleptic attributes such as color [15] or astringency [16] and can, moreover, be used to study wine oxidation [8,17] and ageing [18].…”