基于纳米铕核铁蛋白时间分辨荧光免疫方法的建立

发布时间：2018-04-16 01:05

本文选题：铁蛋白 + 铕　；参考：《现代免疫学》2017年01期

【摘要】：以人铁蛋白(ferritin,FER)为材料,制备纳米铕核微粒,并建立竞争型FER免疫检测方法。用稀盐酸解离FER的亚基后,纯化脱去游离铁,再通过酸碱中和方法重组蛋白外壳,同时将铕离子包裹于去铁蛋白内,制备铕核铁蛋白。经反应条件优化,确定pH 2.0为FER最适解离酸度。FER外壳重组时,铕试剂的添加量约为FER物质量的10 000倍时能获得最大捕获效率。以此建立了铕核-FER时间分辨荧光免疫检测方法,其灵敏度为0.576ng/mL,IC_(50)为13.15ng/mL,批间变异系数CV%15%,非特异性结合率低。用制备成的铕核-FER构建的竞争法TRFIA与FER夹心法试剂盒对照,两者相关系数达0.966。通过上述方法获得免疫活性好、铕含量高的标记用纳米分子,简化了传统的铕标记方法,建立了构建纳米铕核铁蛋白的新方法。
[Abstract]:The europium nanoparticles were prepared by using human ferritin ferrite (ferritin ferrite) as material and a competitive FER immunoassay method was established.After dissociating the subunit of FER with dilute hydrochloric acid, the free iron was purified and removed, then the recombinant protein shell was neutralized by acid and alkali, and the europium ion was encapsulated in the deferritin to prepare europium nuclear ferritin.After optimizing the reaction conditions, it was determined that when pH 2.0 was the optimal dissociation acidity. Fer shell recombination, the maximum capture efficiency could be obtained when the amount of europium reagent was about 10 000 times of the mass of FER.A time-resolved fluorescence immunoassay for europium nucleus-fer was established. The sensitivity of the method was 0.576ng / mL ~ (-1) and the coefficient of variation (CV) was 13.15ng / mL, and the nonspecific binding rate was low.The correlation coefficient between TRFIA and FER sandwich kit was 0.966.A new method for the construction of europium nucleoferritin (NSF) was established by using the above methods to obtain the nanomolecules with good immunological activity and high europium content, which simplified the traditional europium labeling method.
【作者单位】：江苏省原子医学研究所卫生部核医学重点实验室江苏省分子核医学重点实验室;
【基金】：江苏省社会发展基金项目(BE2009631)
【分类号】：R446.6

【参考文献】