Core B: Protein Expression Core

核心 B：蛋白质表达核心

• 批准号: 10710277
• 负责人: Pamela J Bjorkman
• 金额: $ 41.15万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-10 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10710277
• 关键词: Affinity; Affinity Chromatography; Antibodies; Antigens; Baculoviruses; Binding; Biochemical; Biological Assay; Cell Culture Techniques; Cell Line; Cells; Chromatography; Complex; Crystallization; Dedications; Endotoxins; Equipment; Evaluation; Evolution; Gel Chromatography; Goals; HIV; HIV Envelope Protein gp120; HIV envelope protein; HIV-1; Harvest; Healthcare; Immune Targeting; Immune response; Immunization; Immunize; Immunoglobulin G; Incubators; Injections; Insecta; Laboratories; Libraries; Mammalian Cell; Methods; Molecular Sieve Chromatography; Monitor; Monoclonal Antibodies; Mus; Polysaccharides; Procedures; Production; Proteins; Pump; Recombinant Proteins; Resources; Rotation; Saccharomyces cerevisiae; Sterility; Suspensions; System; Temperature; Testing; Yeasts; design; env Gene Products; experimental study; fast protein liquid chromatography; flasks; glycosylation; milligram; monomer; mouse model; neutralizing antibody; programs; protein expression; protein purification; screening; square foot; stable cell line; tissue culture; vaccination strategy

Biochemical, structural, and immunization experiments to evaluate new immunogens designed and selected to induce broadly neutralizing antibodies will require expression and purification of large numbers of proteins, including HIV envelope proteins and antibodies. The Protein Expression Core will provide purified proteins for this program project, which will facilitate the goals of identifying immunogens and defining immunization strategies that elicit a strong and targeted immune response. Antibodies isolated from immunized mice (Project 1) will be expressed and purified for binding and biochemical characterization as well as for structural studies (Project 2). Env proteins, both monomeric gp120s and native-like Env gp140 trimers (SOSIP.664 constructs), will be expressed for immunizations in Project 1, binding studies in Projects 1 and 2, and structural/biochemical analyses in Project 2. Core B will use various expression systems to produce Env proteins with different glycosylation to determine the effects of glycosylation form on the generated immune response. The large number of different antibodies and HIV proteins to be expressed and purified for this program requires a dedicated protein expression core.

生物化学，结构和免疫实验，以评估新的免疫原设计 并被选择以诱导广泛中和抗体的人，将需要表达和纯化 大量的蛋白质，包括HIV包膜蛋白和抗体。蛋白质 表达核心将提供纯化的蛋白质，这将有助于该计划的项目， 目标是确定免疫原和确定免疫策略， 针对性的免疫反应。将从免疫小鼠（项目1）中分离的抗体 表达并纯化用于结合和生物化学表征以及用于结构 研究（项目2）。Env蛋白，单体gp 120和天然样Env gp 140三聚体 （SOSIP.664构建体），将在项目1中表达用于免疫接种， 项目1和2，以及项目2中的结构/生化分析。核心B将使用各种 表达系统，以产生具有不同糖基化的Env蛋白，以确定影响 糖基化形式对产生的免疫反应的影响。大量不同的 抗体和HIV蛋白的表达和纯化需要一个专门的 蛋白表达核心