Despite being known as resistant proteins, peanut allergens (Ara h 1 and Ara h 2) can be digested and cause allergic reactions. Making the allergens more resistant to digestion may aid in non-absorption and excretion ...Despite being known as resistant proteins, peanut allergens (Ara h 1 and Ara h 2) can be digested and cause allergic reactions. Making the allergens more resistant to digestion may aid in non-absorption and excretion of the allergens. Our objectives were to make Ara h 1 and Ara h 2 more resistant to digestion and test them in a model system using trypsin as the digestive enzyme. The resistant allergens were prepared by covalently attaching p-aminobenzamidine (pABA), a protease inhibitor, to peanut allergens in an extract or on a PVDF membrane using glutaraldehyde, and were then tested for resistance to trypsin digestion. SDS-PAGE and Western blot were performed to determine the allergenic capacity of the modified allergens. A control was prepared using glycine instead. Results showed that Ara h 2, when covalently attached with pABA, was more resistant to trypin digestion than the native allergen. Similarly, Ara h 1, prepared on a PVDF membrane and treated with pABA, displayed a resistance to trypsin digestion. Treatment of the allergens with glycine (a control) instead of pABA showed that the modified allergens were as digestible as native allergens. Blot assays showed that the pABA-treated allergens exhibited a lower allergenic capacity than native allergens. It was concluded that pABA, when attached to peanut allergen Ara h 1 or Ara h 2, inhibited digestion of the allergen by trypsin and reduced their allergenic capacity as well.展开更多
1 INTRODUCTIONIn literature,most matrices of affinity chromatography for urokinase(EC 3.4.99.26)purification were prepared by cyanogen bromide activation.However,theseadsorbents usually suffered from the drawback of l...1 INTRODUCTIONIn literature,most matrices of affinity chromatography for urokinase(EC 3.4.99.26)purification were prepared by cyanogen bromide activation.However,theseadsorbents usually suffered from the drawback of leakage of the ligand,particularly inalkaline medium,because of the instability of the isourea linkage between the ligandand the spacer or agarose.Moreover,the positively charged imido group of theN-substituted isourea derivative and the hydrophobicity of the spacers might promotenonspecific adsorption.On the contrary,the adsorbents prepared by the method展开更多
文摘Despite being known as resistant proteins, peanut allergens (Ara h 1 and Ara h 2) can be digested and cause allergic reactions. Making the allergens more resistant to digestion may aid in non-absorption and excretion of the allergens. Our objectives were to make Ara h 1 and Ara h 2 more resistant to digestion and test them in a model system using trypsin as the digestive enzyme. The resistant allergens were prepared by covalently attaching p-aminobenzamidine (pABA), a protease inhibitor, to peanut allergens in an extract or on a PVDF membrane using glutaraldehyde, and were then tested for resistance to trypsin digestion. SDS-PAGE and Western blot were performed to determine the allergenic capacity of the modified allergens. A control was prepared using glycine instead. Results showed that Ara h 2, when covalently attached with pABA, was more resistant to trypin digestion than the native allergen. Similarly, Ara h 1, prepared on a PVDF membrane and treated with pABA, displayed a resistance to trypsin digestion. Treatment of the allergens with glycine (a control) instead of pABA showed that the modified allergens were as digestible as native allergens. Blot assays showed that the pABA-treated allergens exhibited a lower allergenic capacity than native allergens. It was concluded that pABA, when attached to peanut allergen Ara h 1 or Ara h 2, inhibited digestion of the allergen by trypsin and reduced their allergenic capacity as well.
基金Supported by the National Natural Science Foundation of China.
文摘1 INTRODUCTIONIn literature,most matrices of affinity chromatography for urokinase(EC 3.4.99.26)purification were prepared by cyanogen bromide activation.However,theseadsorbents usually suffered from the drawback of leakage of the ligand,particularly inalkaline medium,because of the instability of the isourea linkage between the ligandand the spacer or agarose.Moreover,the positively charged imido group of theN-substituted isourea derivative and the hydrophobicity of the spacers might promotenonspecific adsorption.On the contrary,the adsorbents prepared by the method
