Production of native-like SOSIP trimers and other Env proteins

核心A（三聚体生产）：主要研究者/项目负责人：Klasse，PJ /摩尔，John P. 摘要 三聚体生产核心的目标是支持研究项目的工作， 适量的高质量、切割的可溶性包膜糖蛋白三聚体和相关试剂，以及 将其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人， 阿尔伯特·库波指挥核心的日常活动。 核心小组的任务概述如下。与三聚体生产相关的其他职责 在HIVRAD计划的生命周期内可能会出现相关抗原，并将在 适当 目的1：生产毫克量的高质量SOSIP三聚体和相关Env蛋白 在哺乳动物细胞系中通过瞬时转染。核心会制造出适量的 由项目1设计和评估（在小规模生产运行中），然后选择用于 按比例放大。将与项目1联系，评估三聚体的纯度和抗原质量，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.664三聚体的永久细胞系。核心将使用 目前已建立的制备表达野生型或表位标记的CHO、293 T或293 S细胞系的方法 SOSIP.664三聚体基于各种基因型，在非GMP条件下。 目的3：指导和促进SOSIP三聚体的GMP生产和纯化 条件CMO（Catalent Inc.）将根据GMP标准生产大量BG 505 SOSIP.664三聚体。 一个包括IAVI、BMGF和Scripps CHAVI-ID的联盟的方向。核心将利用其 积累的专业知识，以指导这一进程，包括创造和转让的方法和生产 参考标准三聚体的可实现量。 通过为研究项目和外部合作者提供中央服务，三聚体生产 核心将密切参与整个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.