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）将生产并提供高质量的抗原（GP41-54-GHC的不同变体）和项目1和2的共刺激分子。具体目的是： 目的1。生产高质量的GP41 MPer抗原。该核心的主要责任是为项目1提供足够数量的GP41-mper抗原变体，以制备金纳米颗粒（GNP）和DNA矩阵上的多价抗原，以及在多丙二醇纳米颗粒中的封装。还将向项目2提供重组蛋白，以进行必要的免疫测定以评估B细胞反应（例如ELISA，ELISPOT，ELISPOT，抗原特异性B细胞标记）。根据需要生成新的重组蛋白质构建体。抗原将被表达，纯化，并且在项目1和2中使用之前将仔细监控其质量。 目标2。产生高质量的免疫效应分子。共刺激分子CD154（CD40L）和BLYS将通过与B细胞共同呈现为游离蛋白或复合GNP或DNA基质的B细胞来增强B细胞反应。 CDI 54和BLYS将在项目1中加载到GNP和DNA矩阵上。或者，它们可以直接用作可溶性蛋白（项目2）。在项目1和2中使用之前，将评估蛋白质的结构和功能完整性。