Protein Core will be the provider of all the important reagents for Project 1 and Project 2 of this grant proposal. It will use the information coming from Project 4 to generate HIV Envelope trimers that will be used as baits for identification of novel neutralizing antibodies in Project 1 and 2. The number of baits generated by Protein Core will be 10-20 per patient. The amount will vary depending on the need of the researchers in other projects. The Core has the capacity to produce up to 100 mg of trimers. With the progress of the Program Protein Core will generate neutralizing antibodies identified with the help of these baits. There will be two dozen antibodies generated per patient. Protein Core has experstise to do cell culture necessary to generate all these proteins. The Core has successfully purifed several grams of a broadly neutralizing antibody that was used in a SHIV protection experiment in non-human primates. Protein Core will perform transient transfections and use mammalian cells, HEK 293T and CHO, to produce the proteins, then we will perform affinity chromatography to purify the proteins for further use by other researchers or further characterization. Protein Core will do some initial characterization of the neutralizing antibodies by BIAcore, the technology that allows real time sudy of protein interaction. We will study the affinity of novel antibodies toward various HIV Envelope proteins.
Specific Aims Aim 1. Preparation of Envelope constructs in particular gpl40 trimers that will act as baits for identification of neutralizing antibodies and tools for mapping serum neutralizing activity and antibody characterization.
Aim 2. Expression and purification of novel antibodies in particular neutralizing antibodies identified in Projects land 2.
Aim 3. Biochemical characterization of the novel antibodies.
Neutralizing antibodies are important for a successful HIV vaccine. Protein Core will produce reagents based on the virus Envelope proteins that will serve as baits for identification of these antibodies. After that the Portein Core will produce neutralizing antibodies for further study and characterization by other researchers.
|Karris, Maile Y; Umlauf, Anya; Vaida, Florin et al. (2016) A randomized controlled clinical trial on the impact of CCR5 blockade with maraviroc in early infection on T-cell dynamics. Medicine (Baltimore) 95:e5315|
|Krumm, Stefanie A; Mohammed, Hajer; Le, Khoa M et al. (2016) Mechanisms of escape from the PGT128 family of anti-HIV broadly neutralizing antibodies. Retrovirology 13:8|
|MacLeod, Daniel T; Choi, Nancy M; Briney, Bryan et al. (2016) Early Antibody Lineage Diversification and Independent Limb Maturation Lead to Broad HIV-1 Neutralization Targeting the Env High-Mannose Patch. Immunity 44:1215-26|
|Landais, Elise; Huang, Xiayu; Havenar-Daughton, Colin et al. (2016) Broadly Neutralizing Antibody Responses in a Large Longitudinal Sub-Saharan HIV Primary Infection Cohort. PLoS Pathog 12:e1005369|
|Var, Susanna R; Day, Tyler R C; Vitomirov, Andrej et al. (2016) Mitochondrial injury and cognitive function in HIV infection and methamphetamine use. AIDS 30:839-48|
|Doores, Katie J; Kong, Leopold; Krumm, Stefanie A et al. (2015) Two classes of broadly neutralizing antibodies within a single lineage directed to the high-mannose patch of HIV envelope. J Virol 89:1105-18|
|Shah, Kartik A; Clark, John J; Goods, Brittany A et al. (2015) Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris. Biotechnol Bioeng 112:2624-9|
|Frost, Simon D W; Murrell, Ben; Hossain, A S Md Mukarram et al. (2015) Assigning and visualizing germline genes in antibody repertoires. Philos Trans R Soc Lond B Biol Sci 370:|
|Murrell, Ben; Weaver, Steven; Smith, Martin D et al. (2015) Gene-wide identification of episodic selection. Mol Biol Evol 32:1365-71|
|Ozkumur, Ayca Yalcin; Goods, Brittany A; Love, J Christopher (2015) Development of a High-Throughput Functional Screen Using Nanowell-Assisted Cell Patterning. Small 11:4643-50|
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