Production of native-like SOSIP trimers and other Env proteins

核心A(三聚体制作)：首席调查员/项目总监：Klasse，PJ/Moore，John P. 摘要 三聚体生产核心的目标是通过制定 适量优质、裂解、可溶的包膜糖蛋白三聚体和相关试剂，以及 将它们提供给项目负责人和外部合作者。PJ Klasse博士将担任核心领导，Klasse先生将担任核心领导。 阿尔伯特·库波指导核心的日常活动。 核心的任务概述如下。与生产三聚体和其他物质有关的其他职责 相关抗原可能会在HIVRAD计划的生命周期内出现，并将在 恰如其分。 目的1：生产毫克量高质量的SOSIP三聚体和相关的环境蛋白 在哺乳动物细胞系中通过瞬时转染法。核心将以适当的数量生产， 任何由项目1设计和评估的三聚体(在小规模生产运行中)，然后为 放大。将与项目1联系，对三聚体的纯度和抗原性进行评估，并提供给 项目1和2以及合作者。 目的2：建立和使用表达SOSIP.664三聚体的永久性细胞系。核心将使用 现在已经建立了稳定表达野生型或表位标记的CHO、293T或293S细胞系的方法 SOSIP.664在非GMP条件下，基于各种基因类型的三聚体。 目的3：指导和促进GMP下SOSIP三聚体的生产和纯化 条件。A CMO(Catalent Inc.)将大量生产GMP级BG505 SOSIP.664三聚体 一个还包括IAVI、BMGF和Scripps CHAVI-ID的财团的方向。核心将使用其 积累指导这一进程的专门知识，包括创造和转让方法和生产 可实现的参比标准三聚体数量。 通过为研究项目和外部合作者提供中心服务，Trimer Products CORE将密切参与整个HIVRAD团队的科学计划。

Core A (Trimer Production): Principal Investigator/Program Director: Klasse, PJ / Moore, John P. Abstract The goal of the Trimer Production Core is to support the work of the Research Projects by making appropriate quantities of high quality, cleaved, soluble envelope glycoprotein trimers and related reagents, and providing them to the Project Leaders and external collaborators. Dr. PJ Klasse will be Core Leader, with Mr. Albert Cupo directing the Core's activities on a day-to-day basis. The tasks of the Core are outlined below. Other responsibilities related to the production of trimers and other relevant antigens may arise during the life of the HIVRAD program and will be accomplished as and when appropriate. Aim 1: To produce milligram quantities of high quality SOSIP trimers and related Env proteins in mammalian cell lines by transient transfection. The Core will produce, in appropriate quantities, any trimers that are designed and evaluated (in small-scale production runs) by Project 1, and then selected for scale-up. The trimers will be assessed for purity and antigenic quality, in liaison with Project 1, and supplied to Projects 1 and 2 and collaborators. Aim 2: To create and use permanent cell lines expressing SOSIP.664 trimers. The Core will use now established methods to make stable CHO, 293T or 293S cell lines expressing wild type or epitope-tagged SOSIP.664 trimers based on various genotypes, under non-GMP conditions. Aim 3: To guide and facilitate the production and purification of SOSIP trimers under GMP conditions. A CMO (Catalent Inc.) will make large quantities of GMP-grade BG505 SOSIP.664 trimers under the direction of a consortium that also involves IAVI, BMGF and the Scripps CHAVI-ID. The Core will use its accumulated expertise to guide this process, including the creation and transfer of methods and the production of achievable quantities of reference standard trimers. By providing a central service to both Research Projects, and external collaborators, the Trimer Production Core will be intimately involved in the scientific program of the entire HIVRAD team.