Novel, cost-effective production of immunoaffinity Abs

描述(申请人提供)：科勒和米尔斯坦在1975年发表的开创性论文中提到杂交瘤：“这种培养物可用于医疗和工业用途。”今天，单抗的使用范围从治疗到无数的试剂，但它们的生产模式基本上保持不变，通常涉及某种形式的体外或体内动物细胞培养。虽然这些分子的成本随着时间的推移而下降，但它们的生产成本仍然很高，无论是作为治疗药物还是简单的实验室试剂。目前，用于试剂和诊断的单抗的生产主要有三种方法，包括培养杂交瘤、将杂交瘤重新导入小鼠体内和随后产生腹水以及将特定的重链和轻链抗体基因导入哺乳动物细胞系。最近，我们Rincon制药公司在单细胞、真核、绿藻-莱茵衣藻的叶绿体中表达包括抗体在内的重组蛋白方面取得了相当大的进展。在建立设施的速度、可伸缩性、齿数和资本成本方面，莱茵哈迪尔毛藻的叶绿体比传统的mAb表达平台具有巨大的优势。在这项提议中，我们寻求表达将用作免疫亲和试剂的单抗。我们的合作者，WL麦迪逊的NeoClone，Inc.，已经开发出一种有效的免疫亲和平台，使用E.coliBeta‘表位作为多肽标签，允许一步纯化融合到该标签上的重组表达分子。然而，NeoClone将这项技术商业化的努力取得的成功有限，主要是因为生产用作提纯基质的NT73的成本很高。这一提议的成功结果将证明在莱茵哈迪叶绿体中产生的单抗作为试剂使用的有效性，并最终用于治疗。利用藻类作为单抗的表达平台，无论它们是用作试剂还是治疗药物，都有望降低这些宝贵的生物分子的成本。

DESCRIPTION (provided by applicant): Kohler and Milstein, in their seminal 1975 paper, said of hybridomas: "Such cultures could be valuable for medical and industrial use." Today, the use of monoclonal antibodies (mAbs) ranges from therapeutics to countless reagents, yet their mode of production remains largely unchanged, generally involving some form of in vitro or in vivo animal cell culture. While the cost of these molecules has decreased over time, they remain expensive to produce, whether it be as therapeutics or simple laboratory reagents. Currently, there are three principle means for the production of mAbs for use as reagents and diagnostics, including culture of hybridomas, re-introduction of hybridomas into mice and subsequent generation of ascites and transfection of mammalian cell lines with specific heavy and light chain antibody genes. Recently, we at Rincon Pharmaceuticals, Inc., have made considerable progress in the expression of recombinant proteins, including antibodies, in the chloroplast of the unicellular, eukaryotic, green alga, Chlamydomonas reinhardtii. The chloroplast of C. reinhardtii offers tremendous advantages over traditional mAb expression platforms when it comes to speed, scalability, COGs and capital costs to build a facility. In this proposal, we seek to express mAbs that will be used as immunoaffinity reagents. Our collaborator, NeoClone, Inc. of Madison, Wl, has developed an effective immunoaffinity platform employing the E. coli Beta' epitope as a peptide tag, allowing single step purification of recombinantly expressed molecules fused to this tag. Neoclone's efforts to commercialize this technology, however, have met with limited success, primarily owing to the high cost associated with the production of NT73 for use as the purification matrix. A successful outcome of this proposal would demonstrate the efficacy of mAbs produced in the C. reinhardtii chloroplast for use as reagents, and ultimately therapeutics. Using algae as an expression platform for mAbs, weather they be used as reagents or therapeutics holds great promise for reducing the costs of these valuable bio-molecules.