→ Project Reference: AGL2015-64488-C2-1/2-R

Summary

   The research groups which participate in this coordinated project have been working in the last 10 years in the development of rapid methods for the detection of xenobiotics, a research line capable of generating useful analytical tools for diverse fields, including Food Safety, where we have mainly focused our studies. The exposure of consumers, and especially child population, to toxic substances through diet is a source of concern for citizens. Potentially present in food, water and feed, biotoxins pose a serious problem for human and animal health, causing significant economic losses to the livestock, agriculture and fishery sectors, and can seriously affect the health of drinking water.

   Among the biotoxins with the highest incidence and toxicity, two have strongly called our attention, patulin and anatoxin-a, as there are currently no immunoanalytic methods that allow the determination of these biotoxins in a simple and reliable way. The cause of this striking lack is that it has not yet been possible to obtain high-performance antibodies, probably because they are extremely small molecules with very little structural complexity.

   Given this situation, we have considered for this project the interest of developing immunoanalytical methods for the selective and sensitive determination of these two important biotoxins in those matrices where their presence could constitute a risk. The strong relationship built by the two participating research groups on the generation of useful immunoreactive substances for the determination of chemical contaminants and residues in food has given us the necessary experience and knowledge to face with sufficient guarantees the ambitious challenge contemplated in this project. The high scientific, social and economic impact that could imply the success of this project is an additional motivating factor.

   As a preliminary step to develop bioanalytical systems, it will be necessary to generate the corresponding specific immunoreagents (haptens, conjugates and antibodies). The design and synthesis of functionalized derivatives is a fundamental stage of the project. The generation of suitable bioreceptors depends on the preservation of the physicochemical characteristics of the target biotoxin and on the correct positioning of the spacer arm. Our proposal contemplates the preparation of diverse and stable regioisomeric derivatives of both toxins so that the changes produced by the linker are minimal and the molecule is displayed to the immune system with alternative orientations. The covalent conjugation of the haptens to different proteins will be carried out by coupling methods that were developed in previous projects. We are confident that these strategies will allow us to generate a large collection of high-affinity and specific polyclonal and monoclonal antibodies to each biotoxin. The immunochemical methods included in this project – competitive ELISA, immunochromatographic strips and immunoaffinity columns – are the approximations that we consider most interesting for their particular analytical purposes. The final objective is to validate the applicability of the developed methods for the detection and control of patulin and anatoxin-a in water and food.