Production of native-like SOSIP trimers and other Env proteins

核心B摘要 三聚体生产核心(核心B)的作用是通过制定 适量优质、可溶性包膜糖蛋白三聚体及相关试剂。核心意志 然后将试剂提供给项目负责人和外部合作者，以及要求 接触到我们团队的试剂。核心还将分享其技术诀窍，并提供培训、质量控制和 为团队提供故障排除服务。PJ Klasse博士将成为核心领导人。阿尔伯特·库波先生将指导核心的 在日常活动的基础上。 核心的任务概述如下。与三聚体和三聚体生产有关的其他职责 其他相关试剂可能会在裁决有效期内产生，并将在适当时完成。 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和2设计和评估的三聚体(在小规模生产运行中)，然后选择 用于扩大规模。将对三聚体进行纯度和抗原性评估，然后提供给项目1和2，以 外部合作者和其他要求我们试剂的同事。 目的2：建立和使用表达SOSIP三聚体的永久性细胞系。核心将立即使用 建立稳定表达野生型或表位标记SOSIP的CHO、293T或293S细胞系的方法 以及在非GMP条件下，基于各种基因类型的生殖系靶向三聚体。 目的3：指导和促进GMP下SOSIP三聚体的生产和纯化 条件，并支持临床试验。A CMO(KBI Inc.)已经成功地制造了多克量的 GMP级BG505 SOSIP.664三聚体，由一个财团指导，该财团还涉及IAVI和 BMGF。生产GMP级GT1.1三聚体的第二个翻译计划应于2019年第二季度完成。 核心已经利用其专业知识来指导这两个计划，并将继续为任何新的计划提供指导，包括通过 方法的创建和转让以及可实现数量的参比标准三聚体的生产。 Core还将生产用于临床试验分析计划的SOSIP三聚体。 通过向研究项目和外部合作者提供中心服务，Trimer Production Core将密切参与整个HIVRAD团队的科学计划。

Core B Abstract The role of the Trimer Production Core (Core B) is to support the work of the Research Projects by making appropriate quantities of high quality, soluble envelope glycoprotein trimers and related reagents. The Core will then provide the reagents to the Project Leaders and external collaborators, and to other scientists who request access to our team’s reagents. The Core will also share its knowhow and provide training, quality control and trouble-shooting services to the team. Dr. PJ Klasse will be Core Leader. Mr. Albert Cupo will direct 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 reagents may arise during the life of the award 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 Projects 1 and 2, and then selected for scale-up. The trimers will be assessed for purity and antigenic quality, then supplied to Projects 1 and 2, to external collaborators and to other colleagues who request our reagents. Aim 2: To create and use permanent cell lines expressing SOSIP trimers. The Core will use now established methods to make stable CHO, 293T or 293S cell lines expressing wild type or epitope-tagged SOSIP and germline-targeting 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, and to support clinical trials. A CMO (KBI Inc.) has successfully made multi-gram amounts of GMP-grade BG505 SOSIP.664 trimers under the direction of a consortium that also involves IAVI and the BMGF. A second translational program to produce GMP-grade GT1.1 trimers should be completed in Q2 of 2019. The Core has used its expertise to guide both programs and will continue to do so for any new ones, including by the creation and transfer of methods and the production of achievable quantities of reference standard trimers. The Core will also produce SOSIP trimers for use in clinical trial analytical programs. 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.