The Therapeutically Anti-prion Active Antibody-fragment scFv-W226: Paramagnetic Relaxation-Enhanced NMR Spectroscopy aided Structure Elucidation of the Paratope-epitope Interface
The Therapeutically Anti-prion Active Antibody-fragment scFv-W226: Paramagnetic Relaxation-Enhanced NMR Spectroscopy aided Structure Elucidation of the Paratope-epitope Interface
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治疗性抗朊病毒活性抗体片段 scFv-W226：顺磁弛豫增强核磁共振波谱辅助互补位-表位界面的结构阐明
DOI：
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发表时间：
2010
影响因子：
4.4
通讯作者：
S. Schwarzinger
中科院分区：
文献类型：
--
作者：
C. Mangels;R. Kellner;J. Einsiedel;Philipp R Weiglmeier;P. Rösch;P. Gmeiner;S. Schwarzinger
摘要 抗体已成为不可或缺的试剂，在生物和生物技术分析、诊断以及治疗中有着广泛的应用。在所有情况下，与表位的选择性相互作用至关重要，并且取决于互补位的构象。虽然表位通常以高通量进行绘制，但在相当短的时间内揭示结构见解的方法很少见。我们在这里证明顺磁弛豫增强（PRE）核磁共振波谱是一种强大的工具，可以揭示有关互补决定区域跨越的凹槽中表位取向的结构信息。特别是，我们利用自旋标签 TOAC，它使用标准固相化学与肽表位融合，其特点是与附着在半胱氨酸或赖氨酸残基的侧链功能上的自旋标签相比，其迁移率降低。我们应用该方法来确定朊病毒蛋白螺旋 1 的方向，该螺旋 1 是治疗性抗朊病毒活性 scFv 片段 W226 的表位。
Abstract Antibodies have become indispensable reagents with numerous applications in biological and biotechnical analysis, in diagnostics as well as in therapy. In all cases, selective interaction with an epitope is crucial and depends on the conformation of the paratope. While epitopes are routinely mapped at high throughput, methods revealing structural insights on a rather short timescale are rare. We here demonstrate paramagnetic relaxation-enhanced (PRE) NMR spectroscopy to be a powerful tool unraveling structural information about epitope-orientation in a groove spanned by the complementary determining regions. In particular, we utilize the spin label TOAC, which is fused to the peptidic epitope using standard solid-phase chemistry and which is characterized by a reduced mobility compared to, e.g., spin labels attached to the side-chain functionalities of cysteine or lysine residues. We apply the method to determine the orientation of helix 1 of the prion protein, which is the epitope for the therapeutically anti-prion active scFv fragment W226.
影响因子：
2.9
作者：
Battiste, JL;Wagner, G
通讯作者：
Wagner, G
影响因子：
2.2
作者：
Iwahara, Junji;Tang, Chun;Clore, G. Marius
通讯作者：
Clore, G. Marius