Quail egg ovomucoid can inhibit activation of basophils and eosinophils,while hen egg ovomucoid has been shown to be a major allergen,named Gal d 1.At present,the differences in structure and function between two ovom...Quail egg ovomucoid can inhibit activation of basophils and eosinophils,while hen egg ovomucoid has been shown to be a major allergen,named Gal d 1.At present,the differences in structure and function between two ovomucoid are unclear.We found the homology of ovomucoid in quail eggs and hen eggs reached77%.Compared with hen egg ovomucoid,the distribution of secondary structure was different in AA52-53,AA57-58,AA66-68,AA71-72,AA131-133,AA139-140,AA157-159 and AA184-185.Among 9 epitopes of egg ovomucoid,there were different amino acids from quail egg ovomucoid in 8 epitopes.Recombination quail egg ovomucoid had trypsin inhibition activity and quail egg ovomucoid didn't specifically bind to serum of eggs allergic patients.Quail egg ovomucoid can significantly inhibit RBL-2 H3 cells degranulation and protect cells morphology to a certain extent,indicating quail egg ovomucoid can inhibit cells activation and have potential anti-allergic effects,which is related to trypsin inhibitory activity.The difference in sensitization compare to hen egg ovomucoid may be due to amino acids differences affecting protein structure by changing antigenic epitopes.展开更多
目的建立一种简单的色谱方法以获得保持免疫活性的高纯度卵类粘蛋白。方法将鸡蛋清中的卵粘蛋白等杂蛋白沉淀及pH调节的预处理,处理后蛋清液上样到CM-Sepharose Fast Flow交换层析柱上,然后收集目的蛋白浓缩,通过比较冷冻干燥后分离的...目的建立一种简单的色谱方法以获得保持免疫活性的高纯度卵类粘蛋白。方法将鸡蛋清中的卵粘蛋白等杂蛋白沉淀及pH调节的预处理,处理后蛋清液上样到CM-Sepharose Fast Flow交换层析柱上,然后收集目的蛋白浓缩,通过比较冷冻干燥后分离的卵类粘蛋白的重量与其在全蛋清中的比例来计算其得率,利用电泳分析卵类粘蛋白纯度,采用酶联免疫法(ELISA)检测卵类粘蛋白免疫活性。结果此方法纯化卵类粘蛋白时间短(少于2 h),卵类粘蛋白得率为45.8%,纯度为94.1%。此外,通过该方法纯化的卵类粘蛋白显示出与标准品相似的免疫活性。结论该分离方法操作简单、成本低廉并且能保持卵类粘蛋白的免疫活性,可用于卵类粘蛋白的致敏性研究。展开更多
基金supported by the Beijing Municipal Natural Science Foundation of China(7202100)。
文摘Quail egg ovomucoid can inhibit activation of basophils and eosinophils,while hen egg ovomucoid has been shown to be a major allergen,named Gal d 1.At present,the differences in structure and function between two ovomucoid are unclear.We found the homology of ovomucoid in quail eggs and hen eggs reached77%.Compared with hen egg ovomucoid,the distribution of secondary structure was different in AA52-53,AA57-58,AA66-68,AA71-72,AA131-133,AA139-140,AA157-159 and AA184-185.Among 9 epitopes of egg ovomucoid,there were different amino acids from quail egg ovomucoid in 8 epitopes.Recombination quail egg ovomucoid had trypsin inhibition activity and quail egg ovomucoid didn't specifically bind to serum of eggs allergic patients.Quail egg ovomucoid can significantly inhibit RBL-2 H3 cells degranulation and protect cells morphology to a certain extent,indicating quail egg ovomucoid can inhibit cells activation and have potential anti-allergic effects,which is related to trypsin inhibitory activity.The difference in sensitization compare to hen egg ovomucoid may be due to amino acids differences affecting protein structure by changing antigenic epitopes.
文摘目的建立一种简单的色谱方法以获得保持免疫活性的高纯度卵类粘蛋白。方法将鸡蛋清中的卵粘蛋白等杂蛋白沉淀及pH调节的预处理,处理后蛋清液上样到CM-Sepharose Fast Flow交换层析柱上,然后收集目的蛋白浓缩,通过比较冷冻干燥后分离的卵类粘蛋白的重量与其在全蛋清中的比例来计算其得率,利用电泳分析卵类粘蛋白纯度,采用酶联免疫法(ELISA)检测卵类粘蛋白免疫活性。结果此方法纯化卵类粘蛋白时间短(少于2 h),卵类粘蛋白得率为45.8%,纯度为94.1%。此外,通过该方法纯化的卵类粘蛋白显示出与标准品相似的免疫活性。结论该分离方法操作简单、成本低廉并且能保持卵类粘蛋白的免疫活性,可用于卵类粘蛋白的致敏性研究。
