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High-scale and high-level protein purification

'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:

  • the purification should take place on a scale of 25 up to 50 grams per sample in order to perform in vivo animal studies.
  • purified proteins should be in the native state, requiring mild processing throughout the purification procedure.
  • the samples should have a high purity of more than 90%.
  • the lipopolysaccharide content in the samples should be low, in order to avoid adverse reactions in animals.
  • Protein purification protocol

    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.

    Expertise on purification

    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.

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Dr. Maurits Burgering

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Dr. Maurits Burgering

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