Protein Production Core

蛋白质生产核心

• 批准号: 8042887
• 负责人: Michael W Cho
• 金额: $ 18.23万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8042887
• 关键词: AIDS Vaccines; Antigens; B-Lymphocytes; Complex; DNA; Enzyme-Linked Immunosorbent Assay; Goals; Gold; HIV-1; Immune; Immune response; Immunoassay; Label; Monitor; Polyanhydrides; Preparation; Production; Proteins; Recombinant Proteins; TNFSF5 gene; Variant; base; enzyme linked immunospot assay; nanoparticle; novel vaccines; response; vaccine delivery

The Protein Production Core (Core B) will produce and provide high quality antigens (different variants of gp41-54-GHC) and costimulatory molecules to Projects 1 and 2. The specific aims are: Aim 1. To produce high quality gp41 MPER-based antigens. A major responsibility of this Core is to provide sufficient amount of different variants of gp41-MPER antigens to Project 1 for preparation of multivalent antigens on gold nanoparticles (GNP) and DNA matrix, and for encapsulation in polyanhydride nanoparticles. Recombinant proteins will also be provided to Project 2 for conducting necessary immunoassays to assess B cell responses (e.g. ELISA, ELISPOT, antigen-specific B cell labeling). New recombinant protein constructs will be generated as needed. Antigens will be expressed, purified, and their quality will be carefully monitored prior to usage in Projects 1 and 2. Aim 2. To produce high quality immune effector molecules. Costimulatory molecules CD154 (CD40L) and BLyS will be used to enhance B cell responses by co-presenting them with antigens to B cells either as free proteins or complexed to GNP or DNA matrix. CDI 54 and BLyS will be loaded onto GNP and DNA matrix in Project 1. Alternatively, they can be used directly as soluble proteins (Project 2). Structural and functional integrity of the proteins will be evaluated prior to usage in Projects 1 and 2.

蛋白质生产核心（核心B）将为项目1和项目2生产和提供高质量抗原（gp 41 -54-GHC的不同变体）和共刺激分子。具体目标是： 目标1.生产高质量的gp 41 MPR抗原。该核心的主要职责是为项目1提供足够量的gp 41-MPER抗原的不同变体，用于在金纳米颗粒（GNP）和DNA基质上制备多价抗原，以及用于包封在聚酸酐纳米颗粒中。还将向项目2提供重组蛋白，用于进行必要的免疫测定，以评估B细胞应答（例如ELISA、ELISPOT、抗原特异性B细胞标记）。将根据需要产生新的重组蛋白构建体。抗原将被表达、纯化，并在项目1和2中使用前对其质量进行仔细监测。 目标2.以产生高质量的免疫效应分子。共刺激分子CD 154（CD 40 L）和BLyS将用于增强B细胞应答，通过将它们与抗原以游离蛋白或与GNP或DNA基质复合的形式共呈递给B细胞。CDI 54和BLyS将加载到项目1中的GNP和DNA基质上。或者，它们可以直接用作可溶性蛋白质（项目2）。在用于项目1和2之前，将评价蛋白质的结构和功能完整性。