A differential pulse voltammetric immunoassay associated with the TMB-H2O2-HRP for the determination of IgE antibody was developed. The enzymatic reaction product produced by HRP catalyzing the oxidation of H2O2 with ...A differential pulse voltammetric immunoassay associated with the TMB-H2O2-HRP for the determination of IgE antibody was developed. The enzymatic reaction product produced by HRP catalyzing the oxidation of H2O2 with TMB exhibits a sensitive voltammet-ric response at 0.10V(vs.Ag/AgCl) in pH4.0 phosphate-citrate buffer solution. The de-tection limit of the present method for IgE antibody was 4.0IU/mL, which was four times lower than that obtained by traditional spectrophotometric ELISA procedure. It has been ap-plied to the determination of IgE in human serum with satisfactory results.展开更多
一步法提取鸭血清转铁蛋白是利用热变性沉淀去除鸭血清中的杂蛋白,在微碱性(pH值为8.0)条件下,75℃保温25 m in,转铁蛋白的纯化倍数为1.11,收率为96%,而硫酸铵分级沉淀法的纯化倍数为1.03,收率为51%。一步法与硫酸铵分级沉淀法相比,方...一步法提取鸭血清转铁蛋白是利用热变性沉淀去除鸭血清中的杂蛋白,在微碱性(pH值为8.0)条件下,75℃保温25 m in,转铁蛋白的纯化倍数为1.11,收率为96%,而硫酸铵分级沉淀法的纯化倍数为1.03,收率为51%。一步法与硫酸铵分级沉淀法相比,方法简单,成本低廉,适宜工业化生产。展开更多
实验采用末端快速扩增(RACE)技术从大菱鲆脾脏cDNA文库中筛选得到了免疫球蛋白轻链IgL的全长cD-NA片段。该序列包含47 bp的5′末端非编码区(5′UTR),738 bp的开放阅读框(ORF)和202 bp 3′UTR,整个开放阅读框编码246个氨基酸。系统发生...实验采用末端快速扩增(RACE)技术从大菱鲆脾脏cDNA文库中筛选得到了免疫球蛋白轻链IgL的全长cD-NA片段。该序列包含47 bp的5′末端非编码区(5′UTR),738 bp的开放阅读框(ORF)和202 bp 3′UTR,整个开放阅读框编码246个氨基酸。系统发生分析表明大菱鲆IgL基因与五条鰤的IgL基因起源关系最近。RT-PCR分析表明,大菱鲆IgL基因只在正常脾脏、肾脏和头肾组织中表达;IgL在大菱鲆胚胎发育细胞期就已开始表达,在大菱鲆胚胎尾芽期和体节期表达持续增强;在鳗弧菌感染大菱鲆脾脏和头肾早期均检测到IgL基因强烈表达,后期表达逐渐减弱;大菱鲆胚胎细胞(TEC)在用鳗弧菌感染48h后,IgL开始表达。这些结果均表明IgL基因在大菱鲆免疫应答中起着重要作用。展开更多
文摘A differential pulse voltammetric immunoassay associated with the TMB-H2O2-HRP for the determination of IgE antibody was developed. The enzymatic reaction product produced by HRP catalyzing the oxidation of H2O2 with TMB exhibits a sensitive voltammet-ric response at 0.10V(vs.Ag/AgCl) in pH4.0 phosphate-citrate buffer solution. The de-tection limit of the present method for IgE antibody was 4.0IU/mL, which was four times lower than that obtained by traditional spectrophotometric ELISA procedure. It has been ap-plied to the determination of IgE in human serum with satisfactory results.
文摘实验采用末端快速扩增(RACE)技术从大菱鲆脾脏cDNA文库中筛选得到了免疫球蛋白轻链IgL的全长cD-NA片段。该序列包含47 bp的5′末端非编码区(5′UTR),738 bp的开放阅读框(ORF)和202 bp 3′UTR,整个开放阅读框编码246个氨基酸。系统发生分析表明大菱鲆IgL基因与五条鰤的IgL基因起源关系最近。RT-PCR分析表明,大菱鲆IgL基因只在正常脾脏、肾脏和头肾组织中表达;IgL在大菱鲆胚胎发育细胞期就已开始表达,在大菱鲆胚胎尾芽期和体节期表达持续增强;在鳗弧菌感染大菱鲆脾脏和头肾早期均检测到IgL基因强烈表达,后期表达逐渐减弱;大菱鲆胚胎细胞(TEC)在用鳗弧菌感染48h后,IgL开始表达。这些结果均表明IgL基因在大菱鲆免疫应答中起着重要作用。
