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

项目摘要 这SBIR紧急竞争修订将开发第一个亲和膜纯化治疗mRNA， 高选择性和高通量。基于mRNA的药物有可能解决各种各样的 病理学基于mRNA的疫苗可以提高安全性并大大缩短开发时间， 大流行的情景。许多基于mRNA的COVID-19疫苗正在开发中，其中一种疫苗 疫苗已完成第一阶段临床试验，并显示出应对COVID-19的巨大潜力 流行病然而，一家开创mRNA药物的公司透露， 下游纯化工艺是工业放大中必须解决的主要障碍， mRNA生产以产生必要的数量和质量。考虑到COVID-19的深远影响， 将对全球近70亿人口，时间来开发一个高生产力的mRNA 净化技术，就像现在提出的那样。通过应对这一挑战，提出的技术 将对mRNA的产生产生显著影响，并通过关联，改善患者对药物的可及性。 疫苗治疗性mRNA通常具有聚腺苷酸（poly-A）尾。寡脱氧胸苷（oligo- dT）已被认为是通过以下方式从进料流中分离多聚腺苷酸化mRNA的有效亲和配体 poly-A尾中的腺嘌呤和oligo-dT中的脱氧胸苷之间的杂交。本次竞赛的目标 修订项目是为了证明开发基于dT的亲和膜产品的可行性， 结合能力的快速和选择性纯化的聚腺苷酸化的mRNA。初步数据显示， 取得了令人鼓舞从这一创新中衍生出来的产品将是市场上第一种一次性膜 可以将mRNA纯化生产率提高高达一百倍的层析柱， 与常规树脂柱相比具有更高的纯度和产率。本研究的主要目的是（1） 合成和表征mRNA亲和膜和（2）测试原型亲和膜柱， 多聚腺苷酸化mRNA的捕获步骤纯化。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交联条件对容量和回收率的影响 使用市售的纯化mRNA。在具体目标2中，Purilogics将与合作伙伴合作 合同生产组织，以量化捕获步骤纯化的膜柱性能， 用体外转录（IVT）方法制备的多腺苷酸化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 改善膜性能。新列产品的立即市场进入将销售到 生物制药公司的纯化科学家和工程师。

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.