The POM monoclonals: a comprehensive set of antibodies to non-overlapping prion protein epitopes.
The POM monoclonals: a comprehensive set of antibodies to non-overlapping prion protein epitopes.
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DOI：
10.1371/journal.pone.0003872
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发表时间：
2008
期刊：
影响因子：
3.7
通讯作者：
Aguzzi A
中科院分区：
文献类型：
--
作者：
Polymenidou M;Moos R;Scott M;Sigurdson C;Shi YZ;Yajima B;Hafner-Bratkovic I;Jerala R;Hornemann S;Wuthrich K;Bellon A;Vey M;Garen G;James MN;Kav N;Aguzzi A
PrPSc 是细胞朊病毒蛋白 PrPC 的错误折叠和聚集形式，是引起朊病毒疾病的传播因子的唯一确定的成分。 PrPC 在宿主生物体中的表达对于朊病毒复制和朊病毒神经毒性是必需的。了解朊病毒疾病需要对 PrPC 和 PrPSc 进行详细的结构了解。为了实现这一目标，我们开发了一系列单克隆抗体，称为 POM1 至 POM19，针对小鼠 PrPC 的许多不同表位。三个表位位于 N 端八重复区域内，一个位于中心非结构化区域内，四个表位在 PrPC 的球状 C 近端结构域内不连续。其中一些抗体识别对 PrPSc 中的蛋白酶消化具有弹性的表位。其他抗体可免疫沉淀 PrPC，但不能免疫沉淀 PrPSc。第三组被发现可以免疫沉淀两种 PrP 亚型。后面的一些抗体可以用表位模拟肽封闭，并且与过量的这些肽一起孵育可以对 PrPC 和 PrPSc 进行免疫层析。研究发现近氨基抗体可与重复的 PrPC 表位发生反应，从而大大提高其亲和力。我们还从选定的 POM 中创建了功能性单链微型抗体，尽管其分子量较低，但仍保留了结合特性。因此，POM 系列代表了一套独特的试剂，允许使用多种技术进行研究，包括蛋白质印迹、ELISA、免疫沉淀、构象依赖性免疫测定和基于等离子体表面等离子体共振的测定。
PrPSc, a misfolded and aggregated form of the cellular prion protein PrPC, is the only defined constituent of the transmissible agent causing prion diseases. Expression of PrPC in the host organism is necessary for prion replication and for prion neurotoxicity. Understanding prion diseases necessitates detailed structural insights into PrPC and PrPSc. Towards this goal, we have developed a comprehensive collection of monoclonal antibodies denoted POM1 to POM19 and directed against many different epitopes of mouse PrPC. Three epitopes are located within the N-terminal octarepeat region, one is situated within the central unstructured region, and four epitopes are discontinuous within the globular C-proximal domain of PrPC. Some of these antibodies recognize epitopes that are resilient to protease digestion in PrPSc. Other antibodies immunoprecipitate PrPC, but not PrPSc. A third group was found to immunoprecipitate both PrP isoforms. Some of the latter antibodies could be blocked with epitope-mimicking peptides, and incubation with an excess of these peptides allowed for immunochromatography of PrPC and PrPSc. Amino-proximal antibodies were found to react with repetitive PrPC epitopes, thereby vastly increasing their avidity. We have also created functional single-chain miniantibodies from selected POMs, which retained the binding characteristics despite their low molecular mass. The POM collection, thus, represents a unique set of reagents allowing for studies with a variety of techniques, including western blotting, ELISA, immunoprecipitation, conformation-dependent immunoassays, and plasmon surface plasmon resonance-based assays.
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影响因子：
48
作者：
Sigurdson, Christina J.;Peter, K.;Aguzzi, Adriano
通讯作者：
Aguzzi, Adriano
影响因子：
5.4
作者：
KASCSAK, RJ;RUBENSTEIN, R;DIRINGER, H
通讯作者：
DIRINGER, H
影响因子：
4.1
作者：
Krasemann, S;Jürgens, T;Bodemer, W
通讯作者：
Bodemer, W
影响因子：
2.9
作者：
Lysek, DA;Wüthrich, K
通讯作者：
Wüthrich, K
影响因子：
64.8
作者：
Fischer, MB;Roeckl, C;Aguzzi, A
通讯作者：
Aguzzi, A