Core 2. Protein production and characterization.

项目摘要/摘要 项目名称：科学核心2：蛋白质表达和表征核心 我们将建立并运营一个为合作社服务的蛋白质表达和表征核心 沙眼衣原体纳米支架疫苗研究中心通过提供设施， 专业知识、努力、材料和信息管理，以帮助实现单个项目的目标。 我们将建立和维护生产所需的实验室空间、仪器设备和存储设施， 表达载体、纳米载脂蛋白颗粒和蛋白质的表征、保存和分布 抗原。我们将建立和维护足以培养人才的程序和培训材料 拥有必要的专业知识来支持各个项目的蛋白质生产需求。该设施将 应以下要求配备工作人员，以提供蛋白质生产和表征所需的工作 参与项目。我们将作为按需服务中心运营，尽管常用，但可存档 材料可以批量生产、均分和储存，以便随时分发。正规的生产 对于构成项目疫苗技术基础的易腐烂材料，时间表也可能是必要的， 项目1中用于无细胞翻译的质粒，以及项目2中完全形成的基于RNA的疫苗的NLP， 例如。对于经常使用的材料，我们将建立质量标准并保持质量控制 计划，以确保我们生成的材料结果的重现性。我们将与附属项目合作 汇总生产和表征方案，并确定以下指标的可接受公差 质量控制。我们将建立一个可能是初级的信息管理系统，以便 从核心分发的材料可以追溯到程序、人员、材料和相关 与所有3个项目中分发的材料相关的元数据，如质量评估。其核心是 将高度集中在劳伦斯利弗莫尔国家实验室(LLNL)的人员和设施中，但 LLNL可能无法提供一些关键设备和专业知识，在这种情况下，我们将与外部合作 设施。在时间和资源允许的情况下，蛋白质生产和表征核心将致力于多样化 用于疫苗输送的纳米脂蛋白颗粒组合，NLP的规模化生产，配方较少 易腐烂的NLP制剂，并探索使用单粒子方法，如粒子跟踪，用于 表征和质量控制。蛋白质生产和表征核心也可以作为使用者工作 设施，提供培训和使用可能使个人能够进行研发的设施 在其所在地没有专业知识或设备的项目。

PROJECT SUMMARY/ABSTRACT Project Title: Scientific Core 2: Protein Expression and Characterization Core We will establish and operate a protein expression and characterization core that serves the Cooperative Research Center for Nano-Scaffold Vaccines Against Chlamydia trachomatis (Ct) by providing facilities, expertise, effort, materials, and information management to help achieve the aims of the individual projects. We will establish and maintain facilities with lab space, instrumentation, and storage necessary for production, characterization, preservation, and distribution of expression plasmids, nanolipoprotein particles, and protein antigens. We will establish and maintain procedures and training materials sufficient to develop personnel with the expertise necessary to support the protein production needs of the individual projects. The facility will be staffed to provide the effort required for protein production and characterization, as requested by participating projects. We will operate as an on-demand service center, although commonly used, archivable materials may be produced in bulk, aliquoted, and stored for ready distributions. A regular production schedule may also be necessary for perishable materials that form the basis of a projects' vaccine technology, plasmids for cell-free translation within Project 1, and fully formed NLPs for RNA-based vaccines in project 2, for example. For regularly used materials, we will establish quality metrics and maintain a quality control program to ensure reproducibility of results from materials we generate. We will work with affiliated projects to aggregate production and characterization protocols and to determine acceptable tolerances for metrics of quality control. We will establish an information management system, likely rudimentary to start, such that materials distributed from the core can be traced back to the procedures, personnel, materials, and associated metadata, such as quality assessments, associated with the distributed materials across all 3 Projects. The core will be highly centralized with personnel and facilities at Lawrence Livermore National Laboratory (LLNL), but some key equipment and expertise may not be available at LLNL, in which case we will partner with outside facilities. As time and resources permit, the protein production and characterization core will work to diversify the portfolio of nanolipoprotein particles for vaccine delivery, scale production of NLPs, formulate less perishable preparations of NLPs, and explore use of single particle methods, such as particle tracking, for characterization and quality control. The protein production and characterization core can also work as a user facility, providing training and access to facilities that might enable research and development for individual project that do not have the expertise or equipment at their locations.