Direct sequencing of serum antibodies after infection

感染后血清抗体直接测序

• 批准号: 9253986
• 负责人: Adrian Guthals
• 金额: $ 25.5万
• 依托单位: MAPP BIOPHARMACEUTICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2018-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9253986
• 关键词: Address; Affinity; Affinity Chromatography; Antibodies; Antibody Repertoire; Antigens; B cell repertoire; B-Lymphocytes; Binding; Bioinformatics; Bioterrorism; Blood; Blood Cells; Categories; Cell Line; Cells; Chinese Hamster Ovary Cell; Clone Cells; Collection; Communicable Diseases; Cytomegalovirus; Data; Digestion; Disease; Disease Outbreaks; Ebola Hemorrhagic Fever; Ebola virus; Elements; Emerging Communicable Diseases; Enzyme-Linked Immunosorbent Assay; Epidemic; Epitopes; Filoviridae; Filovirus; Frankfurt-Marburg Syndrome Virus; Generations; Genetic Materials; Glycoproteins; Human; Hybridomas; Immunoglobulin G; Individual; Industry; Infection; Licensure; Methods; Monoclonal Antibodies; Patients; Peripheral; Pharmaceutical Preparations; Phase; Plasma; Plasma Cells; Population; Preparation; Procedures; Proteomics; Recombinants; Research Personnel; Risk; Sampling; Serum; Specificity; Spleen; Sudan; Surface; Survivors; Technology; Testing; Therapeutic; Tissues; Vaccines; Vesicular stomatitis Indiana virus; Virus; Work; Zaire Ebola virus; Zika Virus; antigen binding; antigen challenge; ataxia telangiectasia mutated protein; bone; cross reactivity; deep sequencing; drug development; drug discovery; human disease; human monoclonal antibodies; improved; instrumentation; lymph nodes; mortality; novel; peripheral blood; polyclonal antibody; response; seropositive; single cell sequencing; tandem mass spectrometry

Project Summary Monoclonal antibodies (mAbs) are a well-validated drug platform with exquisite specificity, diversity and potency. They offer the lowest-risk class of drug for development through licensure and offer great potential for addressing emerging and re-emerging infectious diseases. To be prepared as threats, like Ebola (EBOV), Marburg (MARV), and Zika viruses, continue to emerge or re-emerge, rapid discovery capabilities are a critical element. One of the best current methods for discovery of potent human mAbs is isolation of peripheral B-cells from survivors/sero-positive individuals for single cell sequencing or hybridoma generation. However, peripheral B-cells are not always easy to obtain and only represent a small percentage of the total B-cell population across all bodily tissues. We have developed an antibody discovery technology in which only serum antibodies are required, i.e. mAb sequences against a given antigen can be sequenced de novo from polyclonal antibody (pAb) pools without the need for sequencing of genetic material. We propose to further refine this novel proteomic approach and apply it towards discovery of new antibodies in serum obtained from EBOV survivors. Here IgGs from survivor plasma will be purified by filovirus glycoprotein antigen specificity and sequenced independently of B-cells. Given our preliminary data, these efforts may yield mAbs that are cross-reactive to ZEBOV, SUDV, BDBV, and possibly MARV. Viable mAbs will ultimately be developed to become pan-EBOV (ZEBOV, SUDV, BDBV) and/or pan-filovirus (EBOV, MARV) therapeutic products. But perhaps the most significant contribution of this work will be to further develop our rapid antibody discovery approach, which may impact drug discovery in nearly all sectors of the mAb industry, including infectious disease.

项目概要 单克隆抗体 (mAb) 是经过充分验证的药物平台，具有精致的特异性， 多样性和效力。他们通过许可提供风险最低的药物类别进行开发 并为解决新出现和重新出现的传染病提供了巨大潜力。成为 埃博拉病毒 (EBOV)、马尔堡病毒 (MARV) 和寨卡病毒等威胁不断出现 或者重新出现，快速发现能力是一个关键要素。目前最好的之一 发现有效的人类单克隆抗体的方法是从其中分离外周 B 细胞 幸存者/血清阳性个体进行单细胞测序或杂交瘤生成。然而， 外周 B 细胞并不总是容易获得，并且只占全部细胞的一小部分。 所有身体组织的 B 细胞总数。我们已经开发出抗体发现 仅需要血清抗体的技术，即针对给定的单克隆抗体序列 可以从多克隆抗体 (pAb) 池中重新对抗原进行测序，无需 遗传物质的测序。我们建议进一步完善这种新颖的蛋白质组学方法 将其应用于发现从 EBOV 幸存者获得的血清中的新抗体。这里是 IgG 幸存者血浆中的病毒将通过丝状病毒糖蛋白抗原特异性纯化并测序 独立于 B 细胞。根据我们的初步数据，这些努力可能会产生以下单克隆抗体： 与ZEBOV、SUDV、BDBV 和可能的MARV 有交叉反应。可行的单克隆抗体最终将 开发成为泛埃博拉病毒（ZEBOV、SUDV、BDBV）和/或泛丝状病毒（EBOV、MARV） 治疗产品。但也许这项工作最重要的贡献将是进一步 开发我们的快速抗体发现方法，这可能会影响几乎所有领域的药物发现 单克隆抗体行业的各个部门，包括传染病。