Protein Production Core

蛋白质生产核心(核心B)将为项目1和2生产和提供高质量的抗原(gp41-54-GHC的不同变体)和共刺激分子。具体目标是： 目的1.制备高质量的gp41MPER抗原。这个核心的主要职责是向项目1提供足够数量的不同gp41-MPER抗原变体，用于在金纳米颗粒(GNP)和DNA基质上制备多价抗原，并将其包裹在聚酸酐纳米颗粒中。重组蛋白也将被提供给项目2，用于进行必要的免疫分析，以评估B细胞反应(例如，ELISAT、ELISPOT、抗原特异性B细胞标记)。根据需要，将产生新的重组蛋白构建体。在项目1和2中使用抗原之前，将对抗原进行表达、纯化，并仔细监测其质量。 目的2.制备高质量的免疫效应分子。共刺激分子CD154(CD40L)和BLyS将被用来增强B细胞的反应，方法是将它们作为游离蛋白或与GNP或DNA基质络合的抗原共同呈递给B细胞。CDI 54和BLyS将在项目1中加载到GNP和DNA基质上。或者，它们可以直接作为可溶性蛋白质使用(项目2)。在项目1和2中使用之前，将对蛋白质的结构和功能完整性进行评估。

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.