抗人μ链单克隆抗体的制备、鉴定及初步应用
[Preparation, characterization and preliminary application of monoclonal antibody against human mu chain].
作者信息
Yang Jing, Ding Tian-bing, Ren Jun-ping, Song Jian-hua, Ma Wen-yu
机构信息
Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 May;21(3):325-7.
AIM
To prepare monoclonal antibody (mAb) against human mu chain with high titer and establish a capture ELISA for early serological diagnosis of infectious diseases.
METHODS
BALB/c mice were immunized with human IgM. Hybridoma cell line which could stably secret the mAb to human IgM was established by routine cell fusion technique. mAb's characteristics (titer, Ig subclass, specificity and relative affinity) were identified by indirect ELISA and Western blot, respectively. A capture ELISA was established by using purified mAb to capture specific IgM for early diagnosis of Japanese encephalitis.
RESULTS
One hybridoma cell line 2E5 stably secreting mAb against human IgMmu chain was obtained. The titer of ascites of the mAb was 1 x 10(-6) and the Ig subclass was IgG1(kappa). Relative affinity of 2E5 was 1 x 10 (-5). Western blot analysis showed that mAb 2E5 reacted specifically to mu chain. Both sensitivity and specificity of the capture ELISA in detecting specific IgM in Japanese encephalitis patients sera were high.
CONCLUSION
mAb 2E5 against human mu chain was prepared successfully, and a capture ELISA for early serological diagnosis of Japanese encephalitis was set up.
目的
制备高效价抗人μ链单克隆抗体（mAb），并建立用于传染病早期血清学诊断的捕获ELISA法。
方法
用人IgM免疫BALB/c小鼠。采用常规细胞融合技术建立能稳定分泌抗人IgM mAb的杂交瘤细胞系。分别用间接ELISA和Western blot法鉴定mAb的特性（效价、Ig亚类、特异性和相对亲和力）。利用纯化的mAb建立捕获ELISA法，用于早期诊断日本脑炎。
结果
获得1株稳定分泌抗人IgMμ链mAb的杂交瘤细胞系2E5。该mAb腹水效价为1×10⁻⁶，Ig亚类为IgG1（κ）。2E5的相对亲和力为1×10⁻⁵。Western blot分析表明mAb 2E5与μ链特异性反应。捕获ELISA法检测日本脑炎患者血清中特异性IgM的敏感性和特异性均较高。
结论
成功制备了抗人μ链mAb 2E5，并建立了用于日本脑炎早期血清学诊断的捕获ELISA法。
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