Bivalent Llama Single-Domain Antibody Fragments against Tumor Necrosis Factor Have Picomolar Potencies due to Intramolecular Interactions.
Bivalent Llama Single-Domain Antibody Fragments against Tumor Necrosis Factor Have Picomolar Potencies due to Intramolecular Interactions.
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DOI：
10.3389/fimmu.2017.00867
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发表时间：
2017
影响因子：
7.3
通讯作者：
de Haard H
中科院分区：
文献类型：
--
作者：
Beirnaert E;Desmyter A;Spinelli S;Lauwereys M;Aarden L;Dreier T;Loris R;Silence K;Pollet C;Cambillau C;de Haard H
肿瘤坏死因子 (TNF) 是一种参与炎症病理的细胞因子，其活性可以被抗体或捕获分子抑制。在此，产生了源自美洲驼的重链抗体可变重链结构域（VHH，也称为 Nanobodies™），用于二价构建体的工程化，其以皮摩尔效力拮抗 TNF 与其受体的结合。对三种单体 VHH（VHH#1、VHH#2 和 VHH#3）进行了详细表征，发现它们以亚纳摩尔亲和力结合 TNF。 TNF-VHH 复合物的晶体结构表明，VHH#1 和 VHH#2 在 TNF 与其 TNFR 相互作用区域的中心共享相同的表位，而 VHH#3 则结合不同但部分重叠的表位。这些结构合理化了我们用二价构建体获得的结果，其中两个 VHH 通过不同长度的接头偶联。与传统抗体相反，这些二价 Nanobody™ 构建体可以与单个三聚体 TNF 结合，从而与亲和力结合并阻断细胞因子中三个受体结合位点中的两个。与抗原结合的不同模式以及二价构建体的工程化支持了基于 VHH 的高效治疗实体的设计。
The activity of tumor necrosis factor (TNF), a cytokine involved in inflammatory pathologies, can be inhibited by antibodies or trap molecules. Herein, llama-derived variable heavy-chain domains of heavy-chain antibody (VHH, also called Nanobodies™) were generated for the engineering of bivalent constructs, which antagonize the binding of TNF to its receptors with picomolar potencies. Three monomeric VHHs (VHH#1, VHH#2, and VHH#3) were characterized in detail and found to bind TNF with sub-nanomolar affinities. The crystal structures of the TNF–VHH complexes demonstrate that VHH#1 and VHH#2 share the same epitope, at the center of the interaction area of TNF with its TNFRs, while VHH#3 binds to a different, but partially overlapping epitope. These structures rationalize our results obtained with bivalent constructs in which two VHHs were coupled via linkers of different lengths. Contrary to conventional antibodies, these bivalent Nanobody™ constructs can bind to a single trimeric TNF, thus binding with avidity and blocking two of the three receptor binding sites in the cytokine. The different mode of binding to antigen and the engineering into bivalent constructs supports the design of highly potent VHH-based therapeutic entities.
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DOI：
10.1038/nrrheum.2015.169
发表时间：
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期刊：
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影响因子：
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影响因子：
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