• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Molecular Biology >Antiparallel conformation of knob and hole aglycosylated half-antibody homodimers is mediated by a CH2-CH3 hydrophobic interaction
【24h】

Antiparallel conformation of knob and hole aglycosylated half-antibody homodimers is mediated by a CH2-CH3 hydrophobic interaction

机译：CH2-CH3疏水相互作用介导的旋钮和孔非糖基化半抗体同二聚体的反平行构象

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Bispecific antibody and antibody-like molecules are of wide interest as potential therapeutics that can recognize two distinct targets. Among the variety of ways such molecules have been engineered is by creating "knob" and "hole" heterodimerization sites in the CH3 domains of two antibody heavy chains. The molecules produced in this manner maintain their biological activities while differing very little from the native human IgG sequence. To better understand the knob-into-hole interface, the molecular mechanism of heterodimerization, and to engineer Fc domains that could improve the assembly and purity of heterodimeric reaction products, we sought crystal structures of aglycosylated heterodimeric and homodimeric "knob" and "hole" Fc fragments derived from bacterial expression. The structure of the knob-into-hole Fc was determined at 2.64 ?. Except for the sites of mutation, the structure is very similar to that of the native human IgG1 Fc, consistent with a heterodimer interaction kinetic KD of 1 nM. Homodimers of the "knob" and "hole" mutants were also obtained, and their X-ray structures were determined at resolutions 2.5 ? and 2.1 ?, respectively. Both kinds of homodimers adopt a head-to-tail quaternary structure and thus do not contain direct knob/knob or hole/hole CH3 interactions. The head-to-tail arrangement was disfavored by adding site-directed mutations at F241 and F243 in the CH2 domains, leading to increases in both rate and efficiency of bispecific (heterodimer) assembly.
机译：双特异性抗体和抗体样分子作为可以识别两个不同靶标的潜在疗法而引起广泛关注。在设计这种分子的各种方法中，有一种是在两条抗体重链的CH3域中创建“旋钮”和“孔”异二聚位点。以这种方式产生的分子保持其生物学活性，而与天然人IgG序列差异很小。为了更好地了解旋钮-孔界面，异源二聚化的分子机制，并设计可改善异源二聚反应产物的组装和纯度的Fc结构域，我们寻求了无糖基化异源二聚体和同二聚体“旋钮”和“孔”的晶体结构。 Fc片段源自细菌表达。球形孔Fc的结构确定为2.64Ω。除了突变位点，其结构与天然人IgG1 Fc的结构非常相似，符合异源二聚体相互作用动力学KD <1 nM。还获得了“旋钮”和“孔”突变体的同调异构体，并以2.5？3的分辨率测定了它们的X射线结构。和2.1Ω。两种同型二聚体均采用从头到尾的四元结构，因此不包含直接的旋钮/旋钮或孔/孔CH3相互作用。通过在CH2域中的F241和F243处添加定点突变，不利于从头到尾的安排，从而导致双特异性（异二聚体）装配的速率和效率均提高。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号