Molecular and Structural Studies of Antibody Diversity Mechanisms

抗体多样性机制的分子和结构研究

基本信息

批准号：
9982985
负责人：
Vaughn Vasil Smider
金额：
$ 39.94万
依托单位：
APPLIED BIOMEDICAL SCIENCE INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-30 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9982985
关键词：
Active Sites Amino Acids Antibodies Antibody Diversity Antibody Formation Antibody Repertoire Antigen Targeting Antigens Area Binding Biochemical Biological Biological Response Modifiers Cattle Cells Cellular Assay Color Complementarity Determining Regions Complex Cysteine Data Diagnostic Diagnostics Research Disulfides Electron Microscopy Enzyme Activation Enzymes Epitopes Event Fab Immunoglobulins Family Flow Cytometry Gene Mutation Genes Genetic Genetic Structures HIV HIV Antibodies HIV-1 Human Immune response Immunization Immunize Immunoglobulin G Immunoglobulin Somatic Hypermutation In Vitro Infectious Agent Investigational Therapies Ion Channel Label Left Length Light Maps Matrix Metalloproteinases Mediating Medicine Metalloproteases Methods Modeling Molecular Molecular Genetics Molecular Structure Mus Mutagenesis Mutate Mutation Pathway interactions Pattern Play Polysaccharides Process Property Proteins Reagent Recombinants Role Site-Directed Mutagenesis Structure Surface System Tertiary Protein Structure Testing Therapeutic Time V(D)J Recombination Vaccine Design Vaccines Vertebrates Viral Virus activation-induced cytidine deaminase antibody engineering antigen binding antigen challenge base combinatorial disulfide bond electron crystallography in vivo insight knob protein neutralizing antibody novel response therapeutic candidate vaccine response

项目摘要

Abstract Antibodies are extremely important as the primary mediators of the immune response to infectious agents and vaccines, but also as recombinant molecules used as therapeutics, diagnostics, and research reagents. Thus, understanding the mechanisms by which antibodies form, bind, and neutralize their targets is very important in multiple biomedical areas. Most vertebrate antibody repertoires form their diversity through V(D)J recombination, where combinatorial rearrangement of V, D, and J genes form a vast repertoire where the complementarity determining regions (CDRs) form a relatively flat binding surface for interaction with antigen. Remarkably, cows appear to have a different mechanism for generating diversity and binding antigen; cow antibodies have particularly long CDR H3 regions, a subset of which can be over 70 amino acids long, which form -ribbon “stalk” and disulfide-bonded “knob” minidomains that protrude far from the typical antibody surface. Remarkably, antibodies from this repertoire can potently and broadly neutralize HIV, whereas normal human or mouse antibody repertoires cannot. Therefore, they have unique abilities to bind and neutralize particularly challenging antigens. Here we propose studies to understand in molecular detail the genetic mechanisms underlying formation of these antibodies, the binding properties of the stalk and knob minidomains, and the unique potential of these antibodes to bind bivalently and bispecifically. These studies will provide insight into mechanisms of viral neutralization generally, and particularly HIV neutralization. Additionally, given the potential of these antibodies to bind recessed epitopes, we will generate additional antibodies against particularly challenging antigens like enzymatic active sites and ion channels. We will employ mutagenesis, binding, and structural methods, as well as cellular assays to evaluate the unique properties of these antibodies. Understanding this novel class of antibodies in detail could provide fundamental insights needed for effective vaccine design as well as discovery and engineering of antibodies against some of the most challenging targets in biomedicine. Similarly, our unique antibodies discovered and characterized in this proposal could eventually become therapeutic candidates themselves.

抽象的抗体作为免疫反应的主要介质极其重要感染剂和疫苗，也可用作治疗药物的重组分子，诊断和研究试剂，从而了解抗体的机制。形成、结合和中和它们的靶标在多个生物医学领域非常重要。脊椎动物抗体库通过 V(D)J 重组形成多样性，其中 V、D 和 J 基因的组合重排形成了一个巨大的库，其中确定互补区域 (CDR) 形成相对平坦的相互作用结合表面值得注意的是，奶牛似乎具有不同的产生多样性的机制。和结合抗原；牛抗体具有特别长的 CDR H3 区域，其中的一个子集长度可以超过 70 个氨基酸，形成  带“茎”和二硫键“旋钮” 远离典型抗体表面的微型结构域值得注意的是，抗体来自。该指令可以有效且广泛地中和 HIV，而正常人或小鼠因此，它们具有独特的结合和中和能力。在这里，我们提出研究以了解分子细节。这些抗体形成的遗传机制、茎的结合特性和旋钮微型结构域，以及这些抗体二价结合的独特潜力这些研究将深入了解病毒中和机制。一般来说，特别是艾滋病毒中和。抗体结合凹进表位，我们将产生额外的抗体，特别是我们将采用诱变技术来挑战抗原，例如酶活性位点和离子通道。结合和结构方法以及细胞测定来评估独特的性质详细了解这类新型抗体可以提供帮助。有效的疫苗设计以及发现和工程所需的基本见解同样，针对生物医学中一些最具挑战性的目标的抗体。该提案中发现和表征的抗体最终可能成为治疗药物候选人自己。