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刚地弓形虫RH株主要表面抗原1的原核表达和纯化及免疫反应性检测 被引量:5
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作者 司进 汤俊明 +6 位作者 赵松 殷旭仁 徐明 何伟 华万全 梁幼生 朱荫昌 《中华检验医学杂志》 CAS CSCD 北大核心 2004年第2期107-109,共3页
目的 对重组的刚地弓形虫RH株主要表面抗原 1(SAG1)原核表达载体SAG1/pET 32c ,在大肠杆菌中诱导表达产物进行亲和纯化 β免疫反应性检测评价。 方法 接种含SAG1/pET 32c的BL2 1单菌落至LB肉汤中 ,1∶10 0稀释转种后用 1mmol/L终浓度... 目的 对重组的刚地弓形虫RH株主要表面抗原 1(SAG1)原核表达载体SAG1/pET 32c ,在大肠杆菌中诱导表达产物进行亲和纯化 β免疫反应性检测评价。 方法 接种含SAG1/pET 32c的BL2 1单菌落至LB肉汤中 ,1∶10 0稀释转种后用 1mmol/L终浓度的IPTG诱导表达 ,表达产物用Ni2 + 螯合柱亲和纯化 ,并用免疫印迹技术对纯化的融合蛋白进行免疫反应性检测 ,用重组融合蛋白对弓形虫病 5 2份阳性和 4 0份阴性血清进行初步检测。结果 SAG1/pET 32c在原核系统中 ,经诱导表达出约 370 0 0大小的融合蛋白 ,并以包涵体的形式存在 ;通过Ni2 + 亲和柱纯化可获得SAG1重组融合蛋白。免疫印迹试验表明 ,重组SAG1融合蛋白能被弓形虫阳性血清特异性的识别。初步检测表明 ,阳性和阴性符合率分别为 81% (42 /5 2 )和 95 % (38/40 )。结论 SAG1在原核系统获得高效表达 ,亲和纯化的rSAG1具有较强的免疫反应性 ,对弓形虫病的免疫诊断具有潜在应用价值 ,也为人用或兽用的弓形虫病免疫诊断试剂盒研制奠定了基础。 展开更多
关键词 表面抗原1 原核表达 纯化 免疫反应性检测 弓形虫病 免疫诊断
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Induction of Endothelial Cell Apoptosis by Anti-alpha-enolase Antibody 被引量:3
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作者 Hong-bo Yang Wen-jie Zheng +1 位作者 Xuan Zhang Fu-lin Tang 《Chinese Medical Sciences Journal》 CAS CSCD 2011年第3期152-157,共6页
Objective To assess the prevalence of anti-alpha-enolase antibody in systemic autoimmune diseases in Chinese patients and its role in endothelial cell apoptosis.Methods The reactivity of anti-alpha-enolase antibody in... Objective To assess the prevalence of anti-alpha-enolase antibody in systemic autoimmune diseases in Chinese patients and its role in endothelial cell apoptosis.Methods The reactivity of anti-alpha-enolase antibody in a variety of autoimmune disorders in Chinese patients was evaluated by dot blot assay.Endothelial cell apoptosis was investigated by in vitro incubation of endothelial cells with IgG purified from anti-alpha-enolase antibody-positive sera,with or without pre-incubation with recombinant alpha-enolase.Results Anti-alpha-enolase antibody was prevalent in different systemic autoimmune diseases with relatively high reactivity in Chinese patients.In vitro incubation of endothelial cells with IgG containing anti-alpha-enolase antibody induced apoptosis in a time-and dose-dependent manner.Apoptosis was partly inhibited by pre-incubation of the endothelial cells with recombinant alpha-enolase.Conclusions Our data suggest that alpha-enolase is a common auto-antigen recognized by anti-endothelial cell antibodies in connective tissue disease.Interaction between alpha-enolase and its autoantibody plays a role in endothelial cell apoptosis.Changes other than cell killing may contribute to the pathogenesis of endothelial damage and microvascular lesions. 展开更多
关键词 anti-alpha-enolase antibody apoptosis endothelial cell
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