High throughput approach for generating human monoclonal antibodies

产生人单克隆抗体的高通量方法

• 批准号: 8904621
• 负责人: Hiep T Tran
• 金额: $ 22.48万
• 依托单位: ABZYME THERAPEUTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8904621
• 关键词: Affinity; Animals; Antibodies; Antibody Formation; Autoantigens; Binding; Biological Assay; Biological Models; Cell surface; Cells; Cholesterol; DNA; Development; Diagnosis; Diploidy; Engineering; Enzyme-Linked Immunosorbent Assay; Enzymes; Evolution; Fluorescence-Activated Cell Sorting; Galactose; Generations; Genes; Genetic Models; Human; IL2RA gene; Immunization; Immunoassay; Immunoglobulin Somatic Hypermutation; In Vitro; Individual; Interstitial Collagenase; Libraries; Light; Malignant Neoplasms; Mammalian Cell; Measures; Medical; Methodology; Monoclonal Antibodies; National Institute of General Medical Sciences; Nature; Partner in relationship; Petromyzon marinus; Phase; Process; Productivity; Proteins; Reagent; Regulatory T-Lymphocyte; Research; Rheumatoid Arthritis; Saccharomyces cerevisiae; Sensitivity and Specificity; Specificity; Surface; Surface Plasmon Resonance; System; Therapeutic; Therapeutic antibodies; Time; TimeLine; Toxin; Yeasts; activation-induced cytidine deaminase; analog; base; cross reactivity; density; disulfide bond; expression vector; human disease; human monoclonal antibodies; immunogenic; improved; in vivo; innovation; interest; novel; pathogen; public health relevance; response; screening; small molecule; success; therapeutic target

 DESCRIPTION (provided by applicant): Thanks to their exquisite specificity and sensitivity, monoclonal antibodies (Mab) have found a broad application in research, diagnosis and therapy. For treatment of human diseases, fully human antibodies are the most desirable as they are generally not immunogenic and well-tolerated. While there are several existing approaches for generation of antibodies, the processes are still time-consuming, labor-intensive, and unpredictable of success. To simplify and to accelerate the generation of fully human antibodies, a novel non-animal approach for antibody production is proposed. The objective will be achieved by using a Self-Diversifying Human Antibody Library (SHALib) effectively displayed on yeast cell surface. SHALib will provide a diverse array of antibodies in vitro, without the need of animal immunization and potential necessary humanization of the antibody. Starting from a naive antibody library from healthy human donors, antibodies with high-affinity and specificity to various targets will be generated and selected through repeated rounds of systematic evolution by somatic hypermutation, target-specific enrichment and fluorescence-activated cell sorting. Since this is a non-animal system, the platform can generate human antibodies against toxins, pathogens, self-antigens, and the like which are by nature difficult to obtain. As a proof-of-principle, in phase I, we will develop human antibodies to five selected targets of therapeutic importance. The targets are for the treatment of high cholesterol (PCSK9), rheumatoid arthritis (MMP1 and MMP13) and cancer (MMP7 and regulatory T cell protein CD25). In Phase II, the platform will be used to develop human monoclonal antibodies to at least 50 targets that are involved in different human diseases. The platform will allow generating quickly human monoclonal antibodies that are of therapeutic potential.

 描述（由应用程序提供）：感谢它们的确切特异性和敏感性，单克隆抗体（MAB）在研究，诊断和治疗中发现了广泛的应用。为了治疗人类疾病，完全人类抗体是最理想的，因为它们通常不是免疫原性和耐受性良好的。尽管有几种现有的抗体生成方法，但这些过程仍然很耗时，劳动密集型且成功。为了简化和加速完全人类抗体的产生，提出了一种新型的非动物抗体生产方法。该目标将通过使用有效显示在酵母细胞表面上的自变化的人类抗体文库（Shalib）来实现。 Shalib将在体外提供一系列潜水抗体，而无需 动物免疫抑制和抗体的潜在人性化。从健康的人类供体的幼稚抗体文库开始，将通过体细胞超突变，靶标特异性酶和荧光激活的细胞分类来生成高亲和力和对各种靶标的特异性的抗体。由于这是一个非动物系统，因此该平台可以生成针对毒素，病原体，自我抗原等的人类抗体，而这种抗体本质上很难获得。作为原理证明，在第一阶段，我们将开发针对五个特定靶标的五个靶标的人类抗体。目标是治疗高胆固醇（PCSK9），类风湿关节炎（MMP1和MMP13）和癌症（MMP7和调节性T细胞蛋白CD25）。在第二阶段中，该平台将用于开发至少50个涉及不同人类疾病的靶标的人类单克隆抗体。该平台将允许快速产生具有热势的人类单克隆抗体。