Rapid structural characterization of most promising novel mAbs and vaccines

最有前途的新型单克隆抗体和疫苗的快速结构表征

• 批准号: 9057431
• 负责人: Andrew Barrett Ward
• 金额: $ 58.47万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9057431
• 关键词: Antibodies; Antigens; Binding Sites; Complex; Cryoelectron Microscopy; Crystallography; Development; Electron Microscopy; Engineering; Experimental Designs; Generations; Goals; Hemagglutinin; Housing; Influenza; Joints; Methods; Modeling; Negative Staining; Proteins; Research; Research Project Grants; Resolution; Structure; Therapeutic; Therapeutic antibodies; Vaccine Antigen; Vaccine Research; Vaccines; Validation; X-Ray Crystallography; anti-influenza; antibody engineering; base; combat; design; influenzavirus; neutralizing antibody; novel; programs; protein complex; receptor binding; scaffold; screening; structural genomics; therapeutic vaccine; tool

The Structure Core will support all three projects in this U19. The goal of our U19 research program is to use structure-based rational design to combat influenza virus from both the therapeutic antibody and vaccine perspective. These endeavors, targeting the receptor-binding site (RBS) on hemagglutinin (HA), require many rounds of design and experimental validation executed in an iterative manner. Thus, the focus of this Core is three-fold: (i) Naturally occurring antibodies from Project 1 will be studied in complex with natural HA molecules. (ii) to solve structures of HA in complex with engineered antibodies from Project 2 for therapeutic applications, and (iii) to solve structures of HA engineered scaffolds in complex with known broadly neutralizing antibodies (bnAbs) in Project 3 for vaccine applications. We will use both electron microscopy (EM) and x-ray crystallography to accomplish these aims and generate sufficient structural coverage to enable development of anti-influenza solutions. Specifically, we will use electron microscopy (EM) to solve structures of HA in complex with engineered antibodies. EM is uniquely suited to rapidly generate structural information of protein complexes in a high-throughput manner. The information from EM can directly inform further structural and design efforts. We will employ both negative stain and cryo-EM methods to evaluate antibody-HA complexes at moderate and high resolution, respectively. We will also use x-ray crystallography to solve structures of HA and engineered HA scaffolds with antibodies. X-ray crystallography is routinely used to reveal atomic-level details of proteins and their complexes. Using the crystal screening capacity of The Joint Center for Structural Genomics housed at Scripps (Dr. Wilson is the PI), we can screen large numbers of antibodies, antigens, and conjugate complexes. We will use this pipeline to solve atomic-resolution structures of antibody-HA and antibody-scaffold complexes.

Structure Core将支持U19中的所有三个项目。我们U19的目标是 研究计划是使用基于结构的合理设计来对抗流感病毒 从治疗性抗体和疫苗的角度来看。这些努力，目标是 血凝素(HA)上的受体结合位点(RBS)需要多轮设计 以及以迭代方式执行的实验验证。因此，这一次的重点 核心是三个方面：(I)项目1的自然产生的抗体将在#年进行研究 与天然HA分子形成复合体。(Ii)解决羟基磷灰石的复杂结构 来自项目2的用于治疗应用的工程抗体，以及(Iii)解决 具有广为人知中和作用的复合羟基磷灰石工程支架的结构 项目3中用于疫苗应用的抗体(BNAbs)。我们将使用这两个电子 显微镜(EM)和X射线结晶学来实现这些目标并产生 有足够的结构覆盖面，以便能够开发抗流感解决方案。 具体地说，我们将使用电子显微镜(EM)来解决HA在络合物中的结构 用基因工程抗体。EM是唯一适合快速生成结构的 高通量的蛋白质复合体信息。来自EM的信息 可以直接为进一步的结构和设计工作提供参考。我们将使用这两种底片 中、高度抗体-HA复合体的染色和冷冻-EM评价方法 分辨率分别为。我们还将使用x射线结晶学来解决HA的结构 并用抗体构建了HA支架。X射线结晶学通常被用来 揭示蛋白质及其复合体的原子级细节。使用水晶筛网 位于斯克里普斯的结构基因组学联合中心的能力(威尔逊博士 等电点)，我们可以筛选出大量的抗体、抗原和结合物。 我们将使用这条流水线来解决抗体-HA的原子分辨结构和 抗体-支架复合体。