New Technology for High-Resolution Antibody Profiling for SARS-CoV-2

SARS-CoV-2 高分辨率抗体分析新技术

• 批准号: 10481680
• 负责人: Mark Lim
• 金额: $ 97.86万
• 依托单位: AMBERGEN, INC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-19 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10481680
• 关键词: 2019-nCoV; Acute; Address; Analysis of Variance; Antibodies; Antibody Response; Antigens; Binding; Biological Assay; Boston; COVID-19; COVID-19 pandemic; COVID-19 patient; Characteristics; Classification; Cohort Analysis; Collaborations; Coupled; Cross Reactions; Device or Instrument Development; Diagnostic; Dimensions; Disease; Epitopes; Evaluation; Feasibility Studies; Fluorescence; Gold; Human; IgG1; Immobilization; Immune; Immune response; Immunoglobulin Isotypes; Individual; Label; Laboratories; Link; Long COVID; Mass Spectrum Analysis; Measures; Methods; Microbiology; Modeling; Nature; Noise; Outcome; Patients; Persons; Phase; Physics; Physiological; Physiology; Play; Postdoctoral Fellow; ROC Curve; Reagent; Reporting; Research; Research Personnel; Resolution; SARS-CoV-2 B.1.617.2; SARS-CoV-2 antibody; SARS-CoV-2 antigen; SARS-CoV-2 exposure; Sampling; Sensitivity and Specificity; Serology; Serum; Severity of illness; Signal Transduction; Structure; System; Technology; Universities; Vaccination; Vaccines; Validation; Variant; Viral Antigens; Virus; Work; antigen binding; base; breakthrough infection; cohort; design; distinguished professor; effective therapy; glycosylation; improved; mass spectrometric imaging; medical schools; new technology; pandemic disease; pathogen; predictive modeling; professor; prognostic; response; severe COVID-19; success; tool; trait; two-dimensional; vaccine hesitancy; virus infection mechanism

Summary/Abstract The SARS-CoV-2 virus has infected to date 35 million and killed over 600,000 persons in the U.S. alone. Despite initial success of the vaccines, the emergence of increasingly more infectious variants such as the delta variant, coupled with vaccine hesitancy and insufficiently effective therapies, has resulted in a continued and deepening national and world-wide pandemic crisis. Moreover, a significant fraction of COVID-19 patients (~35%), even those who are initially asymptomatic, suffer long-term debilitating effects (“long-COVID”). Recent reports correlate the structure of specific SARS-CoV-2 induced antibodies with potentially lethal proinflammatory responses in acute COVID-19. Studies have also linked the antibody response to long-COVID. Moreover, the nature of the antibody response to vaccination correlates with breakthrough infections. Thus, the ability to rapidly perform high- resolution, highly multiplexed antibody response profiling can provide an essential tool, ultimately leading to more effective diagnostics, prognostics, vaccines and treatments for both acute and long-term disease. However, current antibody profiling methods produce a very limited view of the humoral repertoire. To address this unmet need, AmberGen proposes to further develop in Phase II its mass spectrometric bead-array technology for in-depth immune-profiling, termed PC-BAMS-IP™. This will provide researchers with a new and powerful tool for high- resolution antibody profiling which unlike current technology facilitates 2-dimensions of multiplexing. This is accomplished using arrayed photocleavable mass-tag (PC-MT) encoded beads bearing viral antigens to bind serum antibodies, along with a range of PC-MT encoded probes to simultaneously measure the full breadth of bead-bound antibody types. Mass spectrometry imaging (MSI) of the bead-arrays facilitates the decoding of thousands of different PC-MTs, thereby revealing the full complexity of the antibody response and a means to correlate it with disease severity/outcome. Feasibility studies focused on SARS-CoV-2 demonstrate the ability of PC-BAMS-IP™ to perform simultaneous 2-dimensional antibody profiling of both the Fab traits (antigen binding function) and Fc traits (immune effector function). The proposed 2-year Phase II project will expand on this progress, including: i) design, synthesis and evaluation of 25 plug-and-play PC-MT encoded beads for SARS-CoV-2 antigen immobilization and 25 PC-MT probes to simultaneously query a range of Fc traits of the bead-bound serum antibodies; ii) initial validation of the PC-BAMS-IP™ assay using control and COVID-19 convalescent sera, including comparison to Luminex® xMAP® technology, the existing gold-standard for 1-dimensional multiplex antibody profiling; and iii) demonstrate that PC-BAMS-IP™ can distinguish severe and mild COVID-19. This work will be facilitated by our continued collaboration with leading experts including Prof. Cathy Costello (BU, world- renowned mass spectrometry expert), Dr. Jason Amsden (Duke University, mass spectrometric instrument development), Prof. Rahm Gummuluru (BU, Vice Chair of Microbiology, a leading virologist), and Prof. Plamen Ivanov (BU, Director, Keck Laboratory for Network Physiology, advanced statistical physics).

摘要/摘要 迄今为止，仅在美国，SARS-CoV-2 病毒就已感染 3500 万人，并导致超过 60 万人死亡。尽管 疫苗的初步成功，越来越多的传染性变种的出现，例如 Delta 变种， 加上对疫苗的犹豫和有效疗法的不足，导致了持续和深化的 国家和世界范围内的大流行危机。此外，很大一部分 COVID-19 患者（约 35%），即使是那些 最初无症状的人会遭受长期的衰弱影响（“长期新冠病毒”）。最近的报告将 特定 SARS-CoV-2 诱导抗体的结构在急性期具有潜在致命的促炎反应 新冠肺炎。研究还将抗体反应与长期新冠病毒联系起来。此外，抗体的性质 对疫苗接种的反应与突破性感染相关。因此，能够快速执行高 分辨率、高度多重抗体反应分析可以提供一个重要的工具，最终导致更多 针对急性和长期疾病的有效诊断、预后、疫苗和治疗。然而，目前 抗体分析方法对体液库的了解非常有限。为了解决这一未满足的需求， AmberGen 提议在第二阶段进一步开发其质谱珠阵列技术，以深入研究 免疫分析，称为 PC-BAMS-IP™。这将为研究人员提供一个新的、强大的工具来进行高 分辨率抗体分析与当前技术不同，有利于二维多重分析。这是 使用带有病毒抗原的阵列光裂解质量标签 (PC-MT) 编码珠子来结合血清 抗体，以及一系列 PC-MT 编码探针，可同时测量珠子结合的全范围 抗体类型。珠阵列的质谱成像 (MSI) 有助于解码数千个 不同的 PC-MT，从而揭示抗体反应的全部复杂性以及将其与 疾病严重程度/结果。针对 SARS-CoV-2 的可行性研究证明了 PC-BAMS-IP™ 能够 同时对 Fab 特征（抗原结合功能）和 Fc 进行二维抗体分析 特征（免疫效应功能）。拟议的为期 2 年的第二阶段项目将扩大这一进展，包括： i) 设计、合成和评估 25 个用于 SARS-CoV-2 抗原的即插即用 PC-MT 编码珠子 固定化和 25 个 PC-MT 探针可同时查询珠结合血清的一系列 Fc 特征 抗体； ii) 使用对照和 COVID-19 恢复期血清对 PC-BAMS-IP™ 测定进行初步验证， 包括与 Luminex® xMAP® 技术（一维多重分析的现有黄金标准）的比较 抗体分析； iii) 证明 PC-BAMS-IP™ 可以区分严重和轻度 COVID-19。这部作品 我们与包括 Cathy Costello 教授（BU、世界 著名质谱专家）Jason Amsden博士（杜克大学，质谱仪器 开发）、Rahm Gummuluru 教授（BU、微生物学副主任、领先的病毒学家）和 Plamen 教授 伊万诺夫（BU，凯克网络生理学实验室主任，高级统计物理学）。