High throughput approach for generating human monoclonal antibodies

 描述(申请人提供)：由于其精致的特异性和敏感性，单抗在研究、诊断和治疗中得到了广泛的应用。对于人类疾病的治疗，完全的人类抗体是最可取的，因为它们通常不具有免疫原性和耐受性。虽然有几种现有的产生抗体的方法，但这一过程仍然耗时、劳动密集型，并且无法预测成功与否。为了简化和加速完全人类抗体的产生，提出了一种新的非动物抗体产生方法。这一目标将通过有效地展示在酵母细胞表面的自我多样化的人抗体库(Shalib)来实现。沙利布将在体外提供一系列不同的抗体，而不需要 动物免疫和潜在的必要的抗体人性化。从健康供者的天然抗体库出发，通过体细胞超突变、靶点特异性浓缩和荧光激活细胞分选等多轮系统进化，将产生针对不同靶点的高亲和力和特异性抗体。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，这些毒素、病原体、自身抗原等本质上很难获得。作为一项证明原则，在第一阶段，我们将开发针对五个选定的治疗重要靶点的人类抗体。目标是治疗高胆固醇(PCSK9)、类风湿性关节炎(MMP1和MMP13)和癌症(MMP7和调节性T细胞蛋白CD25)。在第二阶段，该平台将用于开发针对至少50个涉及不同人类疾病的靶点的人类单抗。该平台将允许快速产生具有治疗潜力的人类单抗。

 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.