Production of Antibodies in Algae for Biodefense

在藻类中生产抗体用于生物防御

• 批准号: 6984841
• 负责人: STEPHEN P MAYFIELD
• 金额: $ 40.84万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6984841
• 关键词: algae; biomaterial evaluation; biomimetics; biotechnology; bioterrorism /chemical warfare; biotherapeutic agent; chloroplasts; cytoplasm; gene expression; high throughput technology; human genetic material tag; hybrid antibody; immunoglobulin A; immunoglobulin G; immunologic substance development /preparation; intracellular; monoclonal antibody; recombinant proteins; transfection /expression vector

DESCRIPTION (provided by applicant): Monoclonal antibodies (mAbs) have great potential as biodefense agents, whether in passive immunization, detection or disinfection. At present this potential is limited by the cost and time constraints in producing large quantities of these molecules. The most commonly used systems for monoclonal antibody production rely on transformed Chinese Hamster Ovary cells (CHO) grown in suspension culture. Production in CHO cells is a time-consuming process that is cumbersome and expensive to scale up to production volumes. In response to the need for a rapid and inexpensive system for the large scale production of mAbs, we have begun to develop eukaryotic algae as a production platform for the expression of human antibodies. We have successfully developed transformation vectors that allow for the expression of a variety of recombinant proteins, including scFvs, large single chain, and light and heavy chains of IgAs in the chloroplast of the unicellular green alga, Chlamydomonas reinhardtii. Under this proposal, we will expand upon these successes and demonstrate assembly of two classes of full-length human antibodies from both the chloroplast and nucleo-cytoplasmic compartments. Successful completion of this study should provide the proof of concept for using algae as a rapid means to produce large quantities of recombinant human antibodies. Specifically we propose to: 1) Optimize heterologous protein expression in alga chloroplast. 2) Assemble a Fab and full-length human antibody in algal chloroplast. 3) Express and assemble a Fab and full-length antibody from the nucleo-cytoplasmic compartment. 4) Carry out biochemical and cell based analysis of algal expressed antibodies in comparison with their mammalian expressed counterparts. 5) Scale up for production and purification of algal expressed antibodies.

描述（由申请人提供）：单克隆抗体（mAb）作为生物防御剂具有巨大的潜力，无论是在被动免疫、检测还是消毒方面。目前，这种潜力受到大量生产这些分子的成本和时间限制。最常用的单克隆抗体生产系统依赖于悬浮培养中生长的转化中国仓鼠卵巢细胞 (CHO)。 CHO 细胞生产是一个耗时的过程，扩大生产规模既麻烦又昂贵。为了满足大规模生产单克隆抗体的快速且廉价的系统的需求，我们已经开始开发真核藻类作为人类抗体表达的生产平台。我们已经成功开发了转化载体，可在单细胞绿藻莱茵衣藻的叶绿体中表达多种重组蛋白，包括单链抗体、大单链以及 IgA 的轻链和重链。根据这项提议，我们将扩展这些成功，并展示从叶绿体和核细胞质区室组装两类全长人类抗体。这项研究的成功完成应该为使用藻类作为快速生产大量重组人类抗体的方法提供概念证明。具体来说，我们建议： 1) 优化藻类叶绿体中的异源蛋白表达。 2) 在藻类叶绿体中组装 Fab 和全长人抗体。 3) 从核质室表达并组装 Fab 和全长抗体。 4) 对藻类表达的抗体与其哺乳动物表达的对应物进行比较，进行生化和细胞分析。 5) 扩大藻类表达抗体的生产和纯化。