期刊
JOURNAL OF MOLECULAR BIOLOGY
卷 427, 期 16, 页码 2647-2662出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2015.05.016
关键词
Fab; TLR4; in vitro evolution; crystallography; docking
资金
- Swiss National Science Foundation [316030-128787]
- Ligue Genevoise Contre le Cancer grant [1011]
- Boninchi Foundation
- Schmidheiny Foundation
- University of Geneva
- Swiss National Science Foundation (SNF) [316030_128787] Funding Source: Swiss National Science Foundation (SNF)
Hu 15C1 is a potent anti-human Toll-like receptor 4 (TLR4) neutralizing antibody. To better understand the molecular basis of its biological activity, we used a multidisciplinary approach to generate an accurate model of the Hu 15C1 TLR4 complex. By combining site-directed mutagenesis, in vitro antibody evolution, affinity measurements and X-ray crystallography of Fab fragments, we identified key interactions across the Hu 1501 TLR4 interface. These contact points were used as restraints to predict the structure of the Fab region of Hu 1501 bound to TLR4 using computational molecular docking. This model was further evaluated and validated by additional site-directed mutagenesis studies. The predicted structure of the Hu 15C1 TLR4 complex indicates that the antibody antagonizes the receptor dimerization necessary for its activation. This study exemplifies how iterative cycles of antibody engineering can facilitate the discovery of components of antibody-target interactions. (C) 2015 Elsevier Ltd. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据