Production of Antibodies in Algae for Biodefense

描述（由申请人提供）：单克隆抗体（mab）作为生物防御剂具有很大的潜力，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的转化中国仓鼠卵巢细胞（CHO）。在CHO细胞中生产是一个耗时的过程，要扩大产量既麻烦又昂贵。为了应对大规模生产单克隆抗体的快速和廉价系统的需求，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。我们已经成功地开发了转化载体，允许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scFvs，大单链，轻链和重链IgAs。在此建议下，我们将在这些成功的基础上进行扩展，并演示从叶绿体和核细胞质区室中组装两类全长人抗体。这项研究的成功完成应该为利用藻类作为快速生产大量重组人抗体的手段提供概念证明。具体而言，我们建议：

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.