Transgenic Plant Technology for Producing Therapeutic Recombinant Antibodies

描述（由申请人提供）：迫切需要开发新的、具有成本效益的制造工艺来商业化生产治疗性抗体，以降低高制造费用以及解决有限制造能力的可能性。本申请的长期目标是开发使用转基因植物的新型制造方法，以显著降低前期资本投资和加工成本，缩短产品开发周期，并提供具有上级安全特性的产品。这些目标最好通过利用在第一阶段提案的资助下开发的高产转基因莴苣表达系统来实现。在该第二阶段计划中，将使用产生模型抗体的转基因莴苣来评估工艺的可扩展性和来自大规模中试生产的产品质量。在标准哺乳动物细胞培养物中产生的相同模型抗体将用于所有比较。该模型抗体是一个完全人源化的IgG 1？抗组织因子抗体（hOAT），其目前作为抗癌治疗剂处于临床前开发中。本研究的具体目标是：1）在温室环境中使用水培生产产生200-kg转基因hOAT莴苣生物量; 2）建立可扩展的蛋白质纯化方法以从转基因莴苣产生高度纯化的hOAT;和3）使用体外和体内测试评估植物来源的hOAT的批间一致性，并进一步建立其与哺乳动物细胞的可比性。生产hOAT。使用转基因莴苣表达方法的较低的生产和资本成本、缩短的产品开发周期、更大的灵活性和可扩展性以及改进的安全性为该平台提供了用于制造治疗和诊断抗体产品的显著价值和吸引力。较低的商品成本将使抗体疗法得到更广泛的使用，特别是在经济欠发达的市场。快速发展的转基因莴苣能够产生千克数量的功能性重组抗体，非常适合于生物防御应用，其中需要储存治疗性蛋白质来应对众多且不断变化的威胁。使用hOAT作为模型蛋白将提供原理验证，证明有利的生产经济性并确认抗体功能性/功效。它还可能导致植物来源的抗体进入针对癌症适应症的临床研究。7.该提案的目标是开发一种新颖的、具有成本效益的制造工艺，以使用转基因莴苣商业化生产治疗性抗体。这些产品的较低成本和更好的安全性将使抗体治疗剂能够更广泛地使用，特别是在经济欠发达的市场和用于生物防御目的。本研究中使用的拟南芥产生的模型抗体也可以进入抗癌的临床开发。

DESCRIPTION (provided by applicant): There is an urgent need to develop novel, cost-effective manufacturing processes to produce therapeutic antibodies commercially in order to reduce the high manufacturing expenses as well as to address the potential for limited manufacturing capacity. The long-term objectives of this application are to develop novel manufacturing processes using transgenic plants to significantly lower the up-front capital investment and processing costs, shorten the product development cycle, and provide products with superior safety profiles. These objectives are best achieved by utilizing the high-yield transgenic lettuce expression system developed under the funding of the Phase I proposal. In this Phase II plan, transgenic lettuce producing a model antibody will be used to evaluate the scalability of the process and quality of the product from a larger scale pilot production. The same model antibody produced in standard mammalian cell culture will be used for all of comparisons. This model antibody is a fully humanized IgG1? anti-tissue factor antibody (hOAT) that is currently in pre-clinical development as an anti-cancer therapeutic. The specific aims of this research proposal are to: 1) generate 200-kg of transgenic hOAT lettuce biomass using hydroponic production in a greenhouse setting; 2) establish a scalable protein purification method to generate highly purified hOAT from transgenic lettuce; and 3) evaluate lot-to-lot consistency of the plant-derived hOAT using in-vitro and in-vivo testing and further establish its comparability to mammalian cell-produced hOAT. The lower production and capital costs, shortened product development cycle, greater flexibility and scalability, and improved safety of using transgenic lettuce expression methods provides significant value and appeal to this platform for manufacturing therapeutic and diagnostic antibody products. The lower cost of goods would enable wider spread use of antibody therapeutics, particularly in less economically developed markets. The rapidly develop transgenic lettuce capable of producing kilogram quantities of functional recombinant antibody is well suited for biodefense applications where stockpiles of therapeutic proteins are needed to address numerous and continuously changing threats. The use of hOAT as model protein will provide proof-of-principle, demonstrate favorable production economics and confirm antibody functionality/efficacy. It could also lead to the advance of a plant-derived antibody into clinical studies against cancer indications. 7. Project Narrative The goal of this proposal is to develop a novel, cost-effective manufacturing process to commercially produce therapeutic antibodies using transgenic lettuce. The lower cost of goods and better safety profiles of these products would enable broader use of antibody therapeutics, particularly in less economically developed markets and for biodefense purposes. The lettuce-produced model antibody used in this study could also be advanced into clinical development against cancer.