Prevalence and characteristics of anti-PEG antibodies in humans

人类抗 PEG 抗体的流行率和特征

• 批准号: 8622684
• 负责人: Samuel Lai
• 金额: $ 18.83万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8622684
• 关键词: Acute; Address; Adsorption; Affinity; Amino Acids; Animals; Antibodies; B-Lymphocytes; Binding; Biodistribution; Biological Assay; Blood; Blood Banks; Blood Circulation; Blood specimen; Cell Separation; Characteristics; Clinical; Clone Cells; Core Protein; DNA Sequence; Detergents; Development; Docking; Dose; Doxorubicin Hydrochloride Liposome; Drug Formulations; Effectiveness; Ethylene Glycols; Ethylene Oxide; Exposure to; FDA approved; Fertilizers; Food; Future; Hospitals; Human; Immune response; Immunity; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunology; Injection of therapeutic agent; Kinetics; Light; Liposomes; Liver; Measures; Mediating; Molecular Biology; Molecular Conformation; Monoclonal Antibodies; Mucous body substance; Mus; Nature; Opsonin; Patients; Pilot Projects; Polyethylene Glycols; Polymers; Population; Prevalence; Property; Proteins; Publishing; Rattus; Reporting; Research Personnel; Resources; Sampling; Schedule; Serum; Soaps; Specimen; Structure; Testing; Therapeutic; Therapeutic Uses; Time; Toothpaste; Toxic effect; Vertebral column; Water; Work; arm; base; cost; design; ethylene glycol; flexibility; human monoclonal antibodies; immunogenic; improved; in vivo; insight; macrophage; monocyte; nanomedicine; nanoparticle; next generation; particle; posters; public health relevance; repository; response

Abstract Poly(ethylene glycol) (PEG), due to its ability to resist protein adsorption and reduce RES clearance, has been widely used to extend the circulation times and improve efficacy of protein- and nanoparticle- based therapeutics. However, because of constant exposure to PEG in everyday products including detergents (soap, shampoo, toothpaste, etc.), food products and fertilizers, a substantial proportion of the population has likely developed some degree of anti-PEG immunity. Recent animal studies show that antibodies that specifically recognize PEG can be generated in vivo, leading to accelerated blood clearance of PEGylated therapeutics and nanoparticles and thereby reduced efficacy. Although pre-existing anti-PEG immunity likely compromises the effectiveness of many PEGylated therapeutics, little is understood about anti-PEG immunity in humans. In this proposal, we seek to apply the latest advances in immunology and molecular biology to characterize anti-PEG antibodies in humans. In Aim 1, we will measure the prevalence and concentrations of anti-PEG IgG and IgM among blood samples received at the blood bank at the UNC Hospital. We will also evaluate serum anti-PEG antibody levels in a small number of patients before and at various time points post dosing with PEGylated therapeutics. In Aim 2, we will (A) isolate and immortalize human B cells producing anti-PEG antibodies, (B) characterize their binding affinity to PEG (kon, koff, KD), (C) sequence the DNA of high affinity binding monoclonal antibodies (mAb), and (D) obtain crystal structures to reveal how the amino acid orientation of the Fab binding arm specifically recognizes the ethylene oxide backbone of PEG. In Aim 3, using human mAb from Aim 2 cloned to a mouse IgG backbone, we will evaluate (A) how precise levels of anti-PEG IgG, at concentrations reflecting the range found in humans (from Aim 1), influence the bio- distribution and circulation kinetics of PEGylated particles in mice. (B) We will also determine whether pre- injection of free PEG, at various doses and times prior to injection of PEGylated particles, may restore native distribution and circulation characteristics by saturating serum anti-PEG IgG. These studies will contribute valuable resources for future studies in the field (fully human anti-PEG mAb and corresponding multiplex assays), provide important structural insights into how antibodies can specifically recognize PEG and potentially other polymers, and facilitate the development of next generation PEG-like polymers that can avoid binding by anti-PEG antibodies.

抽象的 聚乙二醇 (PEG) 由于具有抵抗蛋白质吸附和降低 RES 清除率的能力，已被 广泛用于延长基于蛋白质和纳米颗粒的循环时间并提高功效 疗法。然而，由于包括洗涤剂在内的日常产品中不断接触 PEG （肥皂、洗发水、牙膏等）、食品和化肥，相当一部分人口拥有 可能产生了某种程度的抗 PEG 免疫力。最近的动物研究表明，抗体 特异性识别 PEG 可在体内生成，从而加速 PEG 化的血液清除 疗法和纳米颗粒，从而降低疗效。尽管预先存在的抗 PEG 免疫力可能 损害许多聚乙二醇化疗法的有效性，但人们对抗聚乙二醇免疫知之甚少 在人类中。在本提案中，我们寻求应用免疫学和分子生物学的最新进展 表征人类抗 PEG 抗体。在目标 1 中，我们将测量以下物质的流行率和浓度： 北卡罗来纳大学医院血库收到的血液样本中含有抗 PEG IgG 和 IgM。我们也会 评估少数患者术前和术后不同时间点的血清抗 PEG 抗体水平 聚乙二醇化治疗剂的剂量。在目标 2 中，我们将 (A) 分离并永生化人类 B 细胞，产生 抗 PEG 抗体，(B) 表征其与 PEG (kon、koff、KD) 的结合亲和力，(C) 对高位点 DNA 进行测序 亲和结合单克隆抗体 (mAb)，以及 (D) 获得晶体结构以揭示氨基酸如何 Fab 结合臂的方向特异性识别 PEG 的环氧乙烷主链。在目标 3 中， 使用来自 Aim 2 的人单克隆抗体克隆至小鼠 IgG 主链，我们将评估 (A) 抗 PEG IgG，其浓度反映了人类中发现的范围（来自目标 1），影响生物- 聚乙二醇化颗粒在小鼠体内的分布和循环动力学。 (B) 我们还将确定是否预 在注射聚乙二醇化颗粒之前以不同剂量和时间注射游离聚乙二醇，可以恢复天然状态。 通过饱和血清抗 PEG IgG 的分布和循环特征。这些研究将有助于 该领域未来研究的宝贵资源（全人抗 PEG mAb 和相应的多重 分析），为抗体如何特异性识别 PEG 和 潜在的其他聚合物，并促进下一代 PEG 类聚合物的开发，以避免 通过抗 PEG 抗体结合。