Producing Humanized Therapeutics in Avian Egg White

用禽蛋清生产人源化治疗药物

• 批准号: 7172267
• 负责人: Leandro Christmann
• 金额: $ 31.06万
• 依托单位: SYNAGEVA BIOPHARMA CORPORATION
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7172267
• 关键词: Antibodies; Antigen Targeting; Artificial Chromosomes; Binding; Bioreactors; Birds; CTLA4 gene; Cells; Chickens; Chromosomes; Clinical Trials; Communicable Diseases; Complementary DNA; Deposition; Drug Industry; Egg White; Embryo; Exhibits; Gene Transfer Techniques; Generations; Genomics; Genus Capra; Goals; Goat; Human; Internal Ribosome Entry Site; Length; Light; Malignant Neoplasms; Mammalian Cell; Methods; Ovomucin; Pharmacologic Substance; Phase; Production; Proteins; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Solutions; Staging; System; Techniques; Testing; Therapeutic; Therapeutic Uses; TimeLine; Transgenic Animals; Transgenic Organisms; antigen binding; base; cost; expression vector; gene delivery system; genetic analysis; glycosylation; human monoclonal antibodies; improved; retroviral-mediated; transmission process

DESCRIPTION (provided by applicant): This phase II SBIR project will test the feasibility of producing large amounts of human monoclonal antibody into transchromosomic avian egg white. We will use a full length ovomucoid genomic locus expression construct created and tested in phase I, to drive expression of the monoclonal's heavy and light chain cDNA. Using these constructs we produced transgenic hens that expressed human monoclonal antibody into their egg whites. This antibody was purified and analyzed for antigen binding and effector function. The results of this analysis support the conclusion that the transgenic produced antibody exhibits antigen binding and effector function that is comparable to the same antibody produced in a mammalian cell culture. This phase II project will also utilize an improved gene delivery system to generate fully transchromosomic avian hens. We have recently generated transchromosomal founders that exhibit germline transmission of a 60MB artificial chromosome. The offspring of these birds contain single copies of the artificial chromosome in every cell. This represents a breakthrough in avian transgenesis which to this point has been restricted by the limited payload capacity of retroviral-mediated methods. We will utilize this improved avian transgenisis technique to deliver the two ovomucoid-based human monoclonal antibody expression vectors developed in phase I to stage I embryos. Transgenic human monoclonal antibody from GO, G1 and G2 birds will be analyzed for expression level, purified and characterized for bioactivity and glycosylation. We anticipate that the G2 production flock will be able to supply sufficient human monoclonal antibody to proceed to clinical trials.

描述（由申请方提供）：该二期SBIR项目将测试在转染色体禽蛋白色中生产大量人单克隆抗体的可行性。我们将使用在I期中创建和测试的全长卵类粘蛋白基因组基因座表达构建体来驱动单克隆重链和轻链cDNA的表达。使用这些构建体，我们产生了转基因母鸡，表达人类单克隆抗体到他们的蛋清。纯化该抗体并分析其抗原结合和效应子功能。该分析的结果支持转基因产生的抗体表现出与哺乳动物细胞培养物中产生的相同抗体相当的抗原结合和效应子功能的结论。第二阶段项目还将利用改进的基因传递系统来产生完全转染色体的禽母鸡。我们最近产生了转染色体创始人，表现出生殖系传输的60 MB人工染色体。这些鸟的后代在每个细胞中都含有人工染色体的单拷贝。这代表了禽类转基因的突破，到目前为止，禽类转基因一直受到逆转录病毒介导方法的有限有效载荷能力的限制。我们将利用这一改进的禽转基因技术，提供两个卵类粘蛋白为基础的人单克隆抗体表达载体，在第一阶段的第一阶段胚胎。将分析来自GO、G1和G2禽类的转基因人单克隆抗体的表达水平，纯化并表征其生物活性和糖基化。我们预计G2生产群将能够提供足够的人单克隆抗体进行临床试验。