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.
The major goal of this proposal is to develop novel vaccine delivery platforms to enhance B cell immune responses against HIV-1 with a long-term goal of developing a protective AIDS vaccine. This Protein Production Core will produce high quality antigens as well as immune effector molecules to Projects 1 and 2.
| Vela Ramirez, Julia E; Tygrett, Lorraine T; Hao, Jihua et al. (2016) Polyanhydride Nanovaccines Induce Germinal Center B Cell Formation and Sustained Serum Antibody Responses. J Biomed Nanotechnol 12:1303-11 |
| Narasimhan, Balaji; Goodman, Jonathan T; Vela Ramirez, Julia E (2016) Rational Design of Targeted Next-Generation Carriers for Drug and Vaccine Delivery. Annu Rev Biomed Eng 18:25-49 |
| Vela-Ramirez, Julia E; Goodman, Jonathan T; Boggiatto, Paola M et al. (2015) Safety and biocompatibility of carbohydrate-functionalized polyanhydride nanoparticles. AAPS J 17:256-67 |
| Vela Ramirez, J E; Roychoudhury, R; Habte, H H et al. (2014) Carbohydrate-functionalized nanovaccines preserve HIV-1 antigen stability and activate antigen presenting cells. J Biomater Sci Polym Ed 25:1387-406 |
| Carrillo-Conde, Brenda R; Roychoudhury, Rajarshi; Chavez-Santoscoy, Ana V et al. (2012) High-throughput synthesis of carbohydrates and functionalization of polyanhydride nanoparticles. J Vis Exp : |
