'Protein samples with a weight of over 25 grams and a purity of more than 90%'. That is the result of a protein purification project that TNO has performed for five crop seed multinationals, cooperating within the global scientific network International Life Sciences Institute (ILSI). The project has been a successful start of a new database that can be used for the screening of recombinant crops on allergenic properties of proteins.
Within the project, five different proteins have been purified: ara h1 and ara h2 from peanut, b-lactoglobulin from milk, rubisco from spinach and lipoxygenase from soy. The specific requirements of ILSI were:
With its experience on protocol development and its in-house chromatography facilities, TNO has been able to meet these requirements. Within a short period of time, TNO experts have set-up an effective purification protocol based on structure, size and charge of the target proteins. Using large 2.5 l chromatography columns, the requested high purification degree and sample weight have been obtained. The concentration of lipopolysaccharides was kept very low throughout all purification steps, by applying high-standard cleaning procedures. The five partners within the ILSI project - Monsanto, Dow Chemical, Syngenta Seeds, Bayer Crop Sciences and Dupont - have applied the purified proteins in animal trials. The results of these trials have been used as input for a new database on allergenic properties of proteins.
TNO's expertise on protein purification is not only useful in the field of food allergy, but for the development of protein-based pharmaceuticals and identification of unknown proteins, such as new enzymes or peptides.