Direct sequencing of serum antibodies after infection

项目摘要 单克隆抗体（mAb）是一种经过充分验证的药物平台，具有极高的特异性， 多样性和潜力。他们通过许可证提供风险最低的药物开发 并为应对新出现和重新出现的传染病提供了巨大的潜力。是 为埃博拉病毒、马尔堡病毒和寨卡病毒等威胁的不断出现做好准备 或重新出现，快速发现能力是一个关键因素。最好的电流之一 用于发现有效的人mAb的方法是分离外周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，在这方面， 外周B细胞并不总是容易获得的，并且仅占外周血B细胞的一小部分。 所有身体组织中的B细胞总数我们发现了一种抗体 仅需要血清抗体的技术，即针对给定抗体的mAb序列。 抗原可以从多克隆抗体（pAb）池重新测序，而不需要 遗传物质的测序。我们建议进一步完善这种新的蛋白质组学方法， 将其应用于发现从EBOV幸存者获得的血清中的新抗体。这里IgGs 将通过丝状病毒糖蛋白抗原特异性纯化幸存者血浆中的病毒，并进行测序 与B细胞无关。鉴于我们的初步数据，这些努力可能会产生mAb， 与ZEBOV、SUDV、BDBV和可能的MARV交叉反应。有活力的mAb最终将 发展成为泛EBOV（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药物的发现。 单克隆抗体行业，包括传染病。

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.