ANIONIC COPOLYMER/TRIMETHYLAMINE CORE MICROCARRIER

阴离子共聚物/三甲胺核心微载体

• 批准号: 2896003
• 负责人: William J Hillegas
• 金额: $ 37.42万
• 依托单位: SOLOHILL ENGINEERING, INC.
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-10 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2896003
• 关键词: biotechnology; cell line; copolymer; growth media; method development; microcapsule; protein biosynthesis; tertiary amine; tissue /cell culture; vaccine development

Presence of bovine spongiform encephalopathy (BSE) in the food chain in Europe, and the reaction of regulatory agencies, has created a unique opportunity for a new microcarrier, one that is itself free of animal proteins but will function under sera-free conditions. Results of Phase I experiments have clearly demonstrated proof of principle for the copolymer trimethylamine (TMA) microcarrier. Cell culture experiments during Phase I included attachment, spreading and growth using MDCK, MDBK, VERO, CEF, human diploid fibroblast cell lines and a cell line not proposed but of commercial interest in gene therapy, COS-7. All the above cell lines (except human diploid fibroblasts) when cultured under sera- free conditions, attached quickly, spread rapidly and were harvested with ease. The overall goals of Phase Il research are aimed at the near term development of this unique microcarrier. Specific aims of the research proposed include: (l) demonstrate growth of the cell lines at the five liter scale in our bioreactor on TMA microcarriers, (2) assess TMA microcarrier performance by producing vaccine-strain viruses as endpoints (3) identify process engineering variables that determine protein syntheses, (4) assess TMA microcarriers as a support for gene transfection and (5) using TMA microcarriers assess production of extracellular matrix proteins and extracellular matrix-degrading proteolytic enzymes. PROPOSED COMMERCIAL APPLICATIONS: Domestically manufactured SoloHill microcarriers have found applications in the U. S. animal health industry. Swedish microcarriers are used in Europe under sera-containing conditions. But commercial producers of human biologicals that use serum may very likely change to sera-free processes, driven largely by the presence of BSE in Europe. Research has shown that this new TMA microcarrier functions very well with low adherent cells under sera-free conditions. TMA microcarriers may expand market usage in (l) large scale cell culture, and perhaps in the latest (2) cell therapy and (3) gene therapy applications currently in Phase I Clinical Trials.

牛海绵状脑病（BSE）在食物链中的存在 欧洲，以及监管机构的反应，创造了一个独特的 一种新的微载体，一种本身不含动物的微载体， 蛋白质，但将在无血清条件下发挥作用。阶段结果 实验已经清楚地证明了 共聚物三甲胺（TMA）微载体。细胞培养实验 在阶段I期间包括使用MDCK的附着、铺展和生长， MDBK、VERO、CEF、人二倍体成纤维细胞系和非 已提出但具有商业价值的基因疗法，COS-7。所有上述 细胞系（人二倍体成纤维细胞除外）在血清- 自由条件下，附着迅速，传播迅速，收获与 放松。 第二阶段研究的总体目标是在近期内 这一独特的微载体的开发。研究的具体目标 提出的内容包括：（l）证明细胞系在五个细胞系中的生长 在我们的生物反应器中在TMA微载体上以1升规模进行，（2）评估TMA 以生产疫苗株病毒作为终点的微载体性能 (3)确定决定蛋白质的工艺工程变量 合成，（4）评估TMA微载体作为基因载体 转染和（5）使用TMA微载体评估 细胞外基质蛋白和细胞外基质降解 蛋白水解酶 拟议的商业应用： 国内制造的SoloHill微载体已经得到应用 在美国S.动物保健行业。瑞典微载体用于 在含血清条件下的欧洲。但商业生产商 使用血清人类生物制品很可能变为无血清 这主要是由于欧洲存在BSE。研究 表明这种新的TMA微载体功能非常好， 在无血清条件下培养贴壁细胞。TMA微载体可能会扩大 （l）大规模细胞培养中的市场应用，并且可能在最新的 (2)细胞治疗和（3）目前处于I期的基因治疗应用 临床试验