以锯缘青蟹为研究对象,从免疫鉴定、分离纯化、抗体制备和免疫学分析等方面对其主要过敏原进行研究。首先利用过敏者血清的免疫印迹法,确定锯缘青蟹的主要过敏原为分子量约38kD的蛋白。然后通过制备丙酮粉、等电点沉淀、硫酸铵沉淀及加...以锯缘青蟹为研究对象,从免疫鉴定、分离纯化、抗体制备和免疫学分析等方面对其主要过敏原进行研究。首先利用过敏者血清的免疫印迹法,确定锯缘青蟹的主要过敏原为分子量约38kD的蛋白。然后通过制备丙酮粉、等电点沉淀、硫酸铵沉淀及加热处理对分子量为38kD的主要过敏蛋白进行了高度纯化。该蛋白的pI约为4.5,与虾的原肌球蛋白Pena1性质相近,证实了锯缘青蟹的主要过敏原为原肌球蛋白。通过免疫新西兰大白兔,制备了原肌球蛋白的抗血清,采用Protein A Sepharose亲和层析柱对动物抗体进行了纯化。该抗血清效价高,经4×105倍稀释后仍能与抗原进行反应。该抗体与甲壳类动物及软体动物的原肌球蛋白具有较强的免疫交叉反应,可用于食品过敏原检测。展开更多
Mud crab(Scylla paramamosain)is one of the most consumed crabs in China.Generally,it is believed that dead mud crabs are not edible.Herein,we investigated the postmortem metabolite profile changes of the muscle and he...Mud crab(Scylla paramamosain)is one of the most consumed crabs in China.Generally,it is believed that dead mud crabs are not edible.Herein,we investigated the postmortem metabolite profile changes of the muscle and hepatopancreas of mud crab associated with storage method using nuclear magnetic resonance-based metabolomics coupled with multivariate data analysis.Our results showed that postmortem changes of trimethylamine,organic acids(lactate,formate,and succinate),as well as ATP and its breakdown products are closely affected by the storage method.These metabolites can be used as indicators for monitoring loss of crab freshness.Furthermore,dead mud crabs preserved an edible quality at least for 4 h at room temperature,12h on ice,and 16 d at−20℃,strongly suggesting that dead crab consumption quality should be determined by freshness.They shouldn’t be discarded without considering the effects of different storage conditions on their freshness.展开更多
文摘以锯缘青蟹为研究对象,从免疫鉴定、分离纯化、抗体制备和免疫学分析等方面对其主要过敏原进行研究。首先利用过敏者血清的免疫印迹法,确定锯缘青蟹的主要过敏原为分子量约38kD的蛋白。然后通过制备丙酮粉、等电点沉淀、硫酸铵沉淀及加热处理对分子量为38kD的主要过敏蛋白进行了高度纯化。该蛋白的pI约为4.5,与虾的原肌球蛋白Pena1性质相近,证实了锯缘青蟹的主要过敏原为原肌球蛋白。通过免疫新西兰大白兔,制备了原肌球蛋白的抗血清,采用Protein A Sepharose亲和层析柱对动物抗体进行了纯化。该抗血清效价高,经4×105倍稀释后仍能与抗原进行反应。该抗体与甲壳类动物及软体动物的原肌球蛋白具有较强的免疫交叉反应,可用于食品过敏原检测。
基金Financial support from the 2025 Technological Innova-tion for Ningbo(No.2019B10010)China Agriculture Re-search System-CARS48+1 种基金K.C.Wong Magna Fund in Ningbo Universitythe Special research funding from the Marine Biotechnology and Marine Engineering Dis cipline Group in Ningbo University(No.422004582)are greatly acknowledged.
文摘Mud crab(Scylla paramamosain)is one of the most consumed crabs in China.Generally,it is believed that dead mud crabs are not edible.Herein,we investigated the postmortem metabolite profile changes of the muscle and hepatopancreas of mud crab associated with storage method using nuclear magnetic resonance-based metabolomics coupled with multivariate data analysis.Our results showed that postmortem changes of trimethylamine,organic acids(lactate,formate,and succinate),as well as ATP and its breakdown products are closely affected by the storage method.These metabolites can be used as indicators for monitoring loss of crab freshness.Furthermore,dead mud crabs preserved an edible quality at least for 4 h at room temperature,12h on ice,and 16 d at−20℃,strongly suggesting that dead crab consumption quality should be determined by freshness.They shouldn’t be discarded without considering the effects of different storage conditions on their freshness.
