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（由申请人提供）：由于其精致的特异性和灵敏度，单克隆抗体（Mab）已在研究，诊断和治疗中获得广泛应用。对于人类疾病的治疗，全人抗体是最理想的，因为它们通常没有免疫原性并且耐受性良好。虽然有几种现有的方法用于产生抗体，但该过程仍然是耗时的，劳动密集型的，并且无法预测成功。为了简化和加速全人抗体的产生，提出了一种用于抗体产生的新的非动物方法。该目标将通过使用在酵母细胞表面有效展示的自我多样化人抗体库（SHALib）来实现。SHALib将在体外提供多种抗体，而不需要 动物免疫和潜在必要的抗体人源化。从来自健康人类供体的天然抗体文库开始，将通过体细胞超突变、靶特异性富集和荧光激活细胞分选的重复轮系统进化来产生和选择对各种靶具有高亲和力和特异性的抗体。由于这是一个非动物系统，该平台可以产生针对毒素、病原体、自身抗原等的人类抗体，这些抗体在自然界很难获得。作为原理验证，在第一阶段，我们将开发针对五个选定的具有治疗重要性的靶点的人类抗体。这些靶点用于治疗高胆固醇（PCSK 9）、类风湿性关节炎（MMP 1和MMP 13）和癌症（MMP 7和调节性T细胞蛋白CD 25）。在第二阶段，该平台将用于开发针对至少50种与不同人类疾病有关的靶标的人类单克隆抗体。该平台将允许快速产生具有治疗潜力的人单克隆抗体。