Disulfide bond introduction for general stabilization of immunoglobulin heavy-chain variable domains
Disulfide bond introduction for general stabilization of immunoglobulin heavy-chain variable domains
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DOI：
10.1016/j.jmb.2008.01.022
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发表时间：
2008-03-21
影响因子：
5.6
通讯作者：
Muyldermans, Serge
中科院分区：
文献类型：
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作者：
Saerens, Dirk;Conrath, Katja;Muyldermans, Serge
几种抗体片段工程技术旨在增强内在稳定性，但并没有以真正通用的方式应用。在此，提出了一种策略，通过在重链抗体（纳米抗体）的免疫球蛋白重链可变结构域的疏水核心中的两条相对β链之间引入特异性二硫键来实现稳定性的一致获得。除了残基39和87之间的二硫键的合理设计之外，将在残基54和78之间具有额外天然存在的胱氨酸的纳米抗体与不含该胱氨酸的等同纳米抗体进行比较。两种新的二硫键交联以各种组合引入几种纳米抗体中。有趣的是，只有额外天然存在的胱氨酸一致地增加野生型纳米抗体的构象和热稳定性，而不影响抗原结合。(C)2008爱思唯尔有限公司保留所有权利。
Several antibody fragment engineering techniques aim at intrinsic stability enhancement, but are not applied in a truly generic way. Here, a strategy is proposed whereby consistent gain in stability is accomplished by introducing a specific disulfide bond between two opposite beta-strands in the hydrophobic core of the immunoglobulin heavy-chain variable domain of heavy-chain antibodies (Nanobody). Besides the rational design of a disulfide bond between residues 39 and 87, a Nanobody harboring an extra naturally occurring cystine between residues 54 and 78 was compared to an equivalent Nanobody without that cystine. Both novel disulfide cross-links were introduced in several Nanobodies in various combinations. Interestingly, only the extra naturally occurring cystine consistently increased the conformational and thermal stabilities of wild-type Nanobodies without affecting antigen binding. (C) 2008 Elsevier Ltd. All rights reserved.