Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA.
Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA.
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DOI：
10.1016/j.jviromet.2005.03.004
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发表时间：
2005-07
影响因子：
3.1
通讯作者：
Kwang J
中科院分区：
文献类型：
--
作者：
He Q;Du Q;Lau S;Manopo I;Lu L;Chan SW;Fenner BJ;Kwang J
本报告描述了针对 N195 蛋白的几种单克隆抗体的生产，N195 蛋白是 SARS 冠状病毒核衣壳蛋白的主要免疫结构域 [He, Q., Chong, K.H., Chang, H.H., Leung, B., Ling, A.E., Wei, T., Chan, S.W., Ooi, E.E., Kwang, J., 2004。用于检测引起严重急性呼吸综合征的冠状病毒抗体的蛋白质印迹测定法的开发。临床。诊断。实验室。免疫学。 11（2）417-422。]。选择并表征了一种代表性 IgG1 单克隆抗体 (MAb) S-A5D5。 S-A5D5 与 SARS CoV 的重组和天然核衣壳蛋白发生特异性反应。评估了 S-A5D5 与纯化的 N195 蛋白的反应性以及使用 MAb 作为检测抗体来开发抗原捕获 ELISA。抗原捕获 ELISA 可以检测到低至 37.5 pg 的纯化 N 蛋白和 50 TCID50 的 SARS CoV。 MAb S-A5D5 与纯化的 N195 和 SARS CoV 核衣壳抗原的特异性结合被人 SARS 阳性血清和豚鼠抗 N195 血清有效抑制。还可以鉴定 N195 刺突重组杆状病毒感染的 Sf-9 细胞中的 N 蛋白。 SARS确诊患者的18份IFA IgM阳性血清样本中检测到N蛋白，而SARS康复患者的9份样本中未检测到N蛋白。对60份健康个体样本进行检测，未出现假阳性结果；对传染性支气管炎病毒（IBV）、鸡冠状病毒进行检测，未出现交叉反应。因此，这种基于单克隆抗体的抗原捕获 ELISA 是早期诊断 SARS CoV 感染的有力工具。
This report describes the production of several MAbs against N195 protein, a major immunodomain of SARS CoV nucleocapsid protein [He, Q., Chong, K.H., Chang, H.H., Leung, B., Ling, A.E., Wei, T., Chan, S.W., Ooi, E.E., Kwang, J., 2004. Development of a Western blot assay for detection of antibodies against coronavirus causing severe acute respiratory syndrome. Clin. Diagn. Lab. Immunol. 11 (2) 417–422.]. One representative IgG1 monoclonal antibody (MAb), S-A5D5, was selected and characterized. S-A5D5 reacted specifically react with both recombinant and native nucleocapsid protein of SARS CoV. The reactivity of S-A5D5 with purified N195 protein and utilization of the MAb as a detector antibody to develop an antigen capture ELISA was assessed. As little as 37.5 pg of purified N protein and 50 TCID50 of SARS CoV could be detected by the antigen capture ELISA. Specific binding of the MAb S-A5D5 to both purified N195 and SARS CoV nucleocapsid antigen was effectively inhibited by human SARS positive serum and guinea pig anti-N195 serum. The N protein in N195-spike recombinant baculovirus-infected Sf-9 cells could also be identified. N protein was detected in 18 IFA IgM-positive serum samples collected from SARS confirmed patients, but not in nine samples collected from SARS recovery patient. No false positive results were given when 60 samples from healthy individuals were tested, and no cross-reaction occurred when infectious bronchitis virus (IBV), chicken coronavirus, was tested. This monoclonal antibody-based antigen capture ELISA is thus a powerful tool for early diagnosis of SARS CoV infection.
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DOI：
10.1128/cdli.11.2.417-422.2004
发表时间：
2004-03-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
He, QG;Chong, KH;Kwang, J
通讯作者：
Kwang, J
DOI：
10.1128/cdli.12.1.135-140.2005
发表时间：
2005-01-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
Di, B;Hao, W;Che, XY
通讯作者：
Che, XY
影响因子：
9.4
作者：
Lu, LQ;Manopo, I;Kwang, J
通讯作者：
Kwang, J
DOI：
10.1128/cdli.12.2.321-328.2005
发表时间：
2005-02-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
He, Q;Manopo, I;Kwang, J
通讯作者：
Kwang, J
DOI：
10.1016/j.jcv.2003.12.007
发表时间：
2004-07
期刊：
Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
影响因子：
--
作者：
Poon LL;Wong BW;Chan KH;Leung CS;Yuen KY;Guan Y;Peiris JS
通讯作者：
Peiris JS