CPM-2 Removal of Peanut Protein Using Magnetic Nanoparticles for Use in Immunotherapy
Abstract
Peanuts (Arachis hypogaea) are a nutrient dense food source containing large amounts of vitamins, proteins, minerals and fats. Not only are peanuts high in these macronutrients but they provide health benefits such as reducing the risk of cardiovascular disease and diabetes (Bonku 2019). Because of their high nutritional value and low cost, peanuts are consumed in vast amounts throughout the world. However, peanut allergies are the leading cause of food-induced anaphylaxis which can be life-threatening (Mayo 2022). The experiment’s goal is to isolate the most common peanut allergens (Ara h 1-3) from Virginia and Jumbo Runner peanuts to use in immunotherapy for the treatment of peanut allergies. First the peanuts were de-fatted using a 1:1 n-hexane-acetone mix followed by extraction of the proteins using carbonate, TRIS, and borate buffers. Carbonate buffer at pH 10.6 performed the best at extracting allergens into solution. For treatment/isolation of allergens the magnetic particles were coated with chitosan, then phytic or tannic acid and sealed with glutaraldehyde to bond the proteins. Tannic acid-coated particles were especially effective in removing Ara h 1-3. The desorption process was done using 0.1 M NaOH solution to break the bonds between the particles and the adsorbed allergens, producing concentrates of protein for use. Bicinchoninic acid (BCA) assay of Virginia extract treated with tannic acid particles removed an average of 31.67 mg protein/mg particle before desorption. BCA, Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and Fourier transform infrared (FTIR) spectroscopy of desorbed particles showed the removal of some proteins from the particles, especially Ara h 1.
Keywords
Peanut allergens, protein removal, immunotherapy, magnetic nano-particles
CPM-2 Removal of Peanut Protein Using Magnetic Nanoparticles for Use in Immunotherapy
University Readiness Center Greatroom
Peanuts (Arachis hypogaea) are a nutrient dense food source containing large amounts of vitamins, proteins, minerals and fats. Not only are peanuts high in these macronutrients but they provide health benefits such as reducing the risk of cardiovascular disease and diabetes (Bonku 2019). Because of their high nutritional value and low cost, peanuts are consumed in vast amounts throughout the world. However, peanut allergies are the leading cause of food-induced anaphylaxis which can be life-threatening (Mayo 2022). The experiment’s goal is to isolate the most common peanut allergens (Ara h 1-3) from Virginia and Jumbo Runner peanuts to use in immunotherapy for the treatment of peanut allergies. First the peanuts were de-fatted using a 1:1 n-hexane-acetone mix followed by extraction of the proteins using carbonate, TRIS, and borate buffers. Carbonate buffer at pH 10.6 performed the best at extracting allergens into solution. For treatment/isolation of allergens the magnetic particles were coated with chitosan, then phytic or tannic acid and sealed with glutaraldehyde to bond the proteins. Tannic acid-coated particles were especially effective in removing Ara h 1-3. The desorption process was done using 0.1 M NaOH solution to break the bonds between the particles and the adsorbed allergens, producing concentrates of protein for use. Bicinchoninic acid (BCA) assay of Virginia extract treated with tannic acid particles removed an average of 31.67 mg protein/mg particle before desorption. BCA, Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and Fourier transform infrared (FTIR) spectroscopy of desorbed particles showed the removal of some proteins from the particles, especially Ara h 1.