针对具有全长仓鼠朊病毒蛋白的朊病毒蛋白制备单克隆抗体。

Preparation of monoclonal antibodies against prion proteins with full-length hamster PrP.

作者信息

Xiao Xin-Li, Jiang Hui-Ying, Zhang Jin, Han Jun, Nie Kai, Zhou Xiao-Bo, Huang Yin-Xia, Chen Lan, Zhou Wei, Zhang Bao-Yun, Liu Yong, Dong Xiao-Ping

机构信息

School of Medicine, Xi'an Jiao-Tong University, Shaanxi, China.

出版信息

Biomed Environ Sci. 2005 Aug;18(4):273-80.

PMID:16274110

Abstract

OBJECTIVE

To prepare the PrP specific monoclonal antibodies (mAbs) that can be used for the detection of mammalian prions and study of pathogenesis of prion diseases.

METHODS

Several BALB/c mice were immunized with recombinant hamster prion protein (HaPrP). Three hybridoma cell lines designated as B7, B9, and B10, secreting monoclonal antibodies against HaPrP, were established by hybridoma technique. The mAbs reactivities were evaluated with ELISA, Western blot, and immunohistochemistry.

RESULTS

The mAbs produced by these cell lines reacted well with different recombinant hamster PrP proteins. Western blot analyses showed that mAbs B7 and B9 reacted with PrPSc from the scrapie-infected animals after proteinase K digestion with three glycosylated forms. The mAbs exhibited cross-reactivity with various PrPC from several other mammalian species, including humans and cattles. Immunohistochemistry assays confirmed that mAbs B7 and B9 could recognize not only extracellular but also intracellular PrPsSc.

CONCLUSION

The mAbs of prion protein are successfully generated by hybridoma technique and can be applied for the diagnosis of prion associated diseases.

摘要

目的

制备可用于检测哺乳动物朊病毒及研究朊病毒病发病机制的朊蛋白（PrP）特异性单克隆抗体（mAbs）。

方法

用重组仓鼠朊蛋白（HaPrP）免疫数只BALB/c小鼠。通过杂交瘤技术建立了3株分泌抗HaPrP单克隆抗体的杂交瘤细胞系，分别命名为B7、B9和B10。采用酶联免疫吸附测定（ELISA）、蛋白质印迹法（Western blot）和免疫组织化学法评估单克隆抗体的反应活性。

结果

这些细胞系产生的单克隆抗体与不同的重组仓鼠PrP蛋白反应良好。蛋白质印迹分析表明，单克隆抗体B7和B9与经蛋白酶K消化后的3种糖基化形式的羊瘙痒病感染动物的PrPSc发生反应。这些单克隆抗体与包括人类和牛在内的其他几种哺乳动物的各种PrPC表现出交叉反应性。免疫组织化学分析证实，单克隆抗体B7和B9不仅可以识别细胞外的PrPSc，还可以识别细胞内的PrPSc。