Targeting Viral Envelope Glycoproteins with Synthetic Antibodies

用合成抗体靶向病毒包膜糖蛋白

基本信息

批准号：
8688885
负责人：
Jonathan R. Lai
金额：
$ 41.09万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8688885
关键词：
Adopted Affinity Antibodies Antibody Affinity Antibody Formation Antigens Antiviral Agents B cell repertoire Beryllium Binding Biochemical Blocking Antibodies Cell membrane Collaborations Complementarity Determining Regions Crystallization DNA Sequence Rearrangement Development Diagnostics Research Dissection Elements Endosomes Epitopes Exhibits Glycoproteins Goals HIV-1 Humoral Immunities Immunization Immunoglobulin Fragments Immunology Immunotherapeutic agent Immunotherapy Influenza Investigation Libraries Light Membrane Membrane Fusion Membrane Glycoproteins Methods Microbiology Molecular Conformation Monoclonal Antibodies Mutagenesis Oligonucleotides Pathway interactions Peptides Phage Display Play Process Reagent Role Scanning Secondary to Source Specificity Structure Technology Testing Therapeutic Therapeutic Agents Vaccination Vaccine Design Viral Viral Antibodies Viral Antigens Virus Virus Diseases Work antibody engineering base combinatorial cost design influenzavirus insight neutralizing antibody new technology novel novel virus progenitor scaffold therapeutic vaccine tool vaccine development virus development

项目摘要

DESCRIPTION (provided by applicant): Monoclonal antibodies directed against the envelope glycoproteins of membrane viruses such as human immunodeficiency virus type 1 (HIV-1), influenza, and ebolavirus are essential reagents for deciphering mechanisms of viral entry, and identifying epitopes for immunotherapy or vaccine development. Recent work in antibody engineering has demonstrated that specific, high-affinity antibodies can be isolated from simple phage display libraries in which diversity at the antibody complementarity determining regions (CDRs) is encoded by designed, synthetically-derived oligonucleotides ('synthetic antibodies'). Therefore, the synthetic antibody approach circumvents many limitations of traditional antibody isolation methods thereby expanding the scope and specificity with which antigens may be targeted. The overall goal of this proposal is to develop enabling synthetic antibody technologies for applications in the study of viral membrane fusion. In Aim 1, we propose to develop synthetic antibody libraries focused toward viral antigens based on the promiscuous germline segment VH1-69. Many viral antibodies borne from this progenitor exhibit similar modes of interaction with their viral antigens, suggesting that VH1-69 could serve as a scaffold for development of virus- specific synthetic antibody libraries. We will produce VH1-69-based synthetic antibody libraries and screen them against viral targets to evaluate this hypothesis. In Aim 2, we propose to identify conformation-specific antibodies that target the membrane-proximal external region (MPER) of HIV-1 gp41. The MPER is the target of several broadly neutralizing antibodies (NAbs) and therefore the subject of intense investigation for vaccine development. We will identify synthetic antibodies against structurally-constrained MPER peptides, and then characterize neutralization potency of these antibodies to gain insight into the role of MPER epitope conformation on inhibition of membrane fusion. In Aim 3, we propose to isolate synthetic antibodies against the putative fusion intermediates of ebolavirus GP1 and GP2. In comparison to HIV-1 and influenza, relatively few antibodies against GP1 and GP2 have been isolated which has greatly impeded mechanistic understanding of membrane fusion for this virus. Our approach will overcome limitations of other antibody isolation methods that have failed to produce GP1- or GP2-specific antibodies, and yield novel reagents for dissection of fusion intermediates and potential immunotherapeutics.

描述（由申请人提供）：针对膜病毒的包膜糖蛋白（例如人类免疫缺陷病毒1型（HIV-1），流感和埃博氏病）的单克隆抗体是病毒进入机制的基本试剂的必不可少的试剂。抗体工程中的最新研究表明，可以从简单的噬菌体显示库中隔离特定的高亲和力抗体，在抗体互补性确定区域（CDR）的多样性通过设计的，合成的衍生寡核苷酸（合成抗体'）编码。因此，合成抗体方法规定了传统抗体隔离方法的许多局限性，从而扩大了可以靶向抗原的范围和特异性。该提案的总体目标是开发用于在病毒膜融合研究中应用的合成抗体技术。在AIM 1中，我们建议基于滥交种系段VH1-69开发集中于病毒抗原的合成抗体文库。该祖细胞中的许多病毒抗体与病毒抗原具有相似的相互作用模式，这表明VH1-69可以用作发展病毒特异性合成抗体文库的支架。我们将产生基于VH1-69的合成抗体文库，并将其筛选为病毒靶标，以评估这一假设。在AIM 2中，我们建议识别针对HIV-1 GP41膜外部区域（MPER）的构象特异性抗体。 MPER是几种广泛中和抗体（NAB）的靶标，因此是疫苗开发的严格研究主题。我们将鉴定针对结构约束的MPER肽的合成抗体，然后表征这些抗体的中和效力，以深入了解MPER表位构象对抑制膜融合的作用。在AIM 3中，我们建议将合成抗体分离出与埃博拉病毒GP1和GP2的假定融合中间体的分离。与HIV-1和流感相比，已经分离出对GP1和GP2的抗体相对较少，这极大地阻碍了对该病毒膜融合的机械理解。我们的方法将克服未能产生GP1或GP2特异性抗体的其他抗体隔离方法的局限性，并产生用于解剖融合中间体和潜在免疫治疗剂的新型试剂。