SBIR Phase II: Protein A Membrane Columns for Rapid Protein Purification

项目摘要 这一SBIR紧急竞争修改将开发出首个用于纯化治疗性mRNA的亲和膜 高选择性和高产量。以信使核糖核酸为基础的药物有可能解决多种不同的 病理学。基于信使核糖核酸的疫苗可以提高安全性并显著缩短研发时间 大流行情景。一些基于信使核糖核酸的新冠肺炎疫苗正在开发中，其中一种 疫苗已完成I期临床试验，回应新冠肺炎大有可为 大流行。然而，一家开创mRNA药物先河的公司透露，缺乏高产量 下游净化过程是工业生产升级过程中必须解决的主要障碍 信使核糖核酸的产生产生必要的数量和质量。考虑到新冠肺炎的深刻影响 全球将有近70亿人口，有时间开发出高生产力的mrna 净化技术，就像提议的那样，现在就是。通过应对这一挑战，拟议的技术 将对信使核糖核酸的产生产生重大影响，并通过关联改善患者对 疫苗。治疗性信使核糖核酸通常有一个多聚腺苷酸(PolyA)尾巴。寡脱氧胸腺嘧啶核苷 DT)已被认为是从饲料流中分离多腺化mRNA的有效亲和配体 寡核苷酸脱氧胸腺嘧啶核苷与Poly-A尾部腺嘌呤的杂交这场比赛的目标是 修订项目是为了论证开发高亲和力DT基膜产品的可行性 快速、选择性纯化多腺苷化信使核糖核酸的结合能力。初步数据显示 令人鼓舞。从这一创新中衍生出的产品将是市场上第一的一次性薄膜 色谱柱可将信使核糖核酸的提纯效率提高100倍。 与传统的树脂柱相比，纯度和产率更高。研究的具体目的是：(1) 信使核糖核酸亲和性膜的合成与表征及(2)亲和膜柱原型的测试 捕获步骤纯化多腺苷化的mRNA.在具体目标1中，Purilogics将评估所扮演的角色 通过配体结构和密度、合成条件以及结合和洗脱条件对容量和回收率的影响 使用商业上可以买到的纯化的mRNA.在特定的目标2中，Purilogics将与合作伙伴合作 合同制造组织量化膜塔性能，用于捕集步骤净化 用体外转录(IVT)过程制备的多腺苷化的mRNA。原型也将是 以现有产品为基准。合成和性能表征的多次迭代将 提高膜的性能。新专栏产品的直接市场准入将是销售到 生物制药公司的提纯科学家和工程师。

Project Summary This SBIR Urgent Competitive Revision will develop the first affinity membrane to purify therapeutic mRNA with high selectivity and throughput. mRNA-based pharmaceuticals have potential to address a wide variety of pathologies. mRNA-based vaccines can increase safety and dramatically shorten development timelines in pandemic scenarios. A number of mRNA-based COVID-19 vaccines are under development, and one such vaccine has completed its Phase-I clinical trial and showed great promise as a response to the COVID-19 pandemic. However, a company pioneering mRNA medicines has revealed that the lack of high throughput downstream purification processes is a major hurdle that must be addressed in the upscaling of industrial mRNA production to yield the necessary quantity and quality. Considering the profound impact that COVID-19 will have on the global population of nearly seven billion people, the time to develop a high productivity mRNA purification technology, like the one proposed, is now. By addressing this challenge, the proposed technology will have a significant impact on mRNA production and, by association, improve patient accessibility to the vaccine. Therapeutic mRNA usually possesses a polyadenylic acid (poly-A) tail. Oligo-deoxythymidine (oligo- dT) has been recognized as effective affinity ligand to isolate polyadenylated mRNA from feed streams via hybridization between adenine in the poly-A tail and deoxythymidine in oligo-dT. The goal of this Competitive Revision project is to demonstrate the feasibility of developing dT-based affinity membrane products with high binding capacity for the rapid and selective purification of polyadenylated mRNA. Preliminary data are highly encouraging. The products derived from this innovation will be first-in-market, disposable membrane chromatography columns that can improve the mRNA purification productivity up to one hundred times with high purity and yield compared to conventional resin columns. The Specific Aims of the study are to (1) synthesize and characterize mRNA affinity membranes and (2) test prototype affinity membrane columns for capture step purification of polyadenylated mRNA. In Specific Aim 1, Purilogics will evaluate the roles played by ligand structure and density, synthesis conditions, and bind-and-elute conditions on capacity and recovery using a commercially available purified mRNA. In Specific Aim 2, Purilogics will collaborate with a partner contract manufacturing organization to quantify membrane column performance for capture step purification of polyadenylated mRNA prepared with in vitro transcription (IVT) processes. The prototypes also will be benchmarked against existing products. Multiple iterations of synthesis and performance characterization will improve membrane performance. Immediate market entry for the new column products will be sales to purification scientists and engineers in biopharmaceutical companies.