Generation of Monoclonal Antibodies against Human Prion Proteins in PrP0/0 Mice
Generation of Monoclonal Antibodies against Human Prion Proteins in PrP0/0 Mice
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在 PrP0/0 小鼠中产生抗人朊病毒蛋白的单克隆抗体
DOI：
10.1007/bf03401656
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发表时间：
1996
影响因子：
5.7
通讯作者：
W. Bodemer
中科院分区：
文献类型：
--
作者：
S. Krasemann;M. Groschup;S. Harmeyer;G. Hunsmann;W. Bodemer
研究背景朊病毒病（Prion diseases）是一组影响人类和动物的神经退行性疾病。人类疾病包括库鲁病、克雅氏病（CJD）、Gerstmann-Sträussler-Scheinker综合征（GSS）和致死性家族性失眠症（FFI）。朊病毒疾病的致病机制尚未被理解。单克隆抗体提供了有价值的工具，在诊断中，以及在基础研究中，几种疾病，然而，单特异性抗血清或单克隆抗体（mAb）对人朊病毒蛋白，直到现在，还没有提供。材料和方法我们已经开发了一种免疫方案的基础上，核酸注射到非耐受性PrP 0/0小鼠。将编码不同人类朊病毒蛋白的DNA或RNA注射到肌肉组织中，所述朊病毒蛋白包括与CJD、GSS和FFI相关的突变序列。用编码朊病毒蛋白（PRNP）基因的DNA质粒初次接种小鼠，并用DNA、RNA或表达PRNP的重组塞姆利基森林病毒颗粒加强免疫。结果获得了抗人朊蛋白的单克隆抗体，并通过肽酶联免疫吸附试验分析了它们的结合行为。免疫印迹、免疫荧光和免疫沉淀。还测定了它们与来自其他物种的朊病毒蛋白的交叉反应性。我们的单克隆抗体是针对四个不同的线性表位，也可以认识到不连续的地区的原生朊病毒protein.ConclusionsThese抗体应该让我们能够解决有关问题的性质的朊病毒蛋白，以及启动和进展的朊病毒疾病。此外，这些单克隆抗体现在可用于人类和动物朊病毒疾病的诊断。
BackgroundPrion diseases belong to a group of neurodegenerative disorders affecting humans and animals. The human diseases indude kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome (GSS), and fatal familial insomnia (FFI). The pathogenic mechanisms of the prion diseases are not yet understood. Monoclonal antibodies provide valuable tools in the diagnosis, as well as in the basic research, of several diseases; however, monospecific antisera or monoclonal antibodies (mAbs) against human prion proteins were, until now, not available.Materials and MethodsWe have developed an immunization protocol based on nucleic acid injection into nontolerant PrP0/0 mice. DNA or RNA coding for different human prion proteins including the mutated sequences associated with CJD, GSS, and FFI were injected into muscle tissue. Mice were primarily inoculated with DNA plasmids encoding the prion protein (PRNP) gene and boosted either with DNA, RNA, or recombinant Semliki Forest Virus particles expressing PRNP. Hybridomas were then prepared.ResultsDifferent mAbs against human prion proteins were obtained, and their binding behavior was analyzed by peptide enzyme-linked immunosorbent assay. Western blot, immunofluorescence, and immunoprecipita-tion. Their cross-reactivity with prion protein from other species was also determined. Our mAbs are directed against four different linear epitopes and may also recognize discontinuous regions of the native prion protein.ConclusionsThese antibodies should allow us to address questions concerning the nature of the prion protein as well as the initiation and progression of prion diseases. Moreover, these mAbs can now be used for the diagnosis of prion diseases of humans and animals.
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DOI：
10.1073/pnas.83.8.2310
发表时间：
1986-04-01
影响因子：
11.1
作者：
MEYER, RK;MCKINLEY, MP;PRUSINER, SB
通讯作者：
PRUSINER, SB
DOI：
10.1073/pnas.85.18.6617
发表时间：
1988-09-01
影响因子：
11.1
作者：
GABIZON, R;MCKINLEY, MP;PRUSINER, SB
通讯作者：
PRUSINER, SB
朊病毒脂质体。
DOI：
10.1042/bj2660001
发表时间：
1990
期刊：
The Biochemical journal
影响因子：
--
作者：
Gabizon,R;Prusiner,SB
通讯作者：
Prusiner,SB
使用合成肽的单特异性抗血清表征朊病毒蛋白。
DOI：
--
发表时间：
1988
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Barry,RA;Vincent,MT;Kent,SB;Hood,LE;Prusiner,SB
通讯作者：
Prusiner,SB
影响因子：
3.7
作者：
XIANG, ZQ;SPITALNIK, S;ERTL, HCJ
通讯作者：
ERTL, HCJ