→ Project Reference: RTI2018-096121-B-C21
Consumer exposure, particularly children, to toxic chemical substances through the diet constitutes an issue of major concern for citizens. Among the contaminants potentially present in food and feed, mycotoxins represent a major problem for human and animal health, causing important economic costs to the agro-food industry. For this group of contaminants, antibody-based methods are widely implemented in laboratories worldwide as screening techniques.
This proposal focuses in the development of immunoanalytical techniques enabling the simple and reliable detection of three of the most relevant mycotoxins in food and feed, that is, zearalenone, alternariol, and patulin. The reasons for the selection of these mycotoxins are diverse. Concerning patulin, it is worth mentioning that, to date, it has not been possible to obtain useful antibodies for the development of rapid methods, most likely because patulin is a very small and low structurally complex molecule. Regarding alternariol, commercial immunoanalytical tests are not available either, although some research groups have made some progress to reach this goal. Nevertheless, the strategies employed so far were essentially based on a single chemical approach, which, in our opinion, is not the most adequate. The same reasoning can be applied to zearalenone, yet in this case the available immunoreagents are sensitive-enough as to allow their implementation in commercial assays. Our long-standing experience in this research field lets us hypothesize that there are alternative, non-explored strategies that should reasonably give rise to antibodies and derived techniques with higher detectability than those developed so far.
As a previous step to the development of the new bioanalytical systems, the generation of the corresponding specific immunoreagents (haptens, bioconjugates, and antibodies) will be necessary. Preserving the physico-chemical properties of the mycotoxin and choosing the correct derivatization site in haptens will be the key factors for the generation of antibodies with the desired properties. Our proposal encompasses the preparation of several regioisomeric derivatives of each toxin with different linker tethering sites, in a way that minimum modifications are introduced and the hapten can be displayed to the immune system in alternative orientations. We are confident that these strategies will culminate with the generation of a wide collection of high-affinity and specific monoclonal antibodies to zearalenone, alternariol, and patulin. Each one of the immunoanalytical techniques included in this proposal, competitive ELISA, immunochromatographic strips, and immunoaffinity columns, will serve for a different analytical demand. Ultimately, the applicability of the new techniques will be validated in a range of products, including fruits, juices, cereals, and feed.