Core B Abstract The role of the Trimer Production Core (Core B) is to support the work of the Research Projects by making appropriate quantities of high quality, soluble envelope glycoprotein trimers and related reagents. The Core will then provide the reagents to the Project Leaders and external collaborators, and to other scientists who request access to our team?s reagents. The Core will also share its knowhow and provide training, quality control and trouble-shooting services to the team. Dr. PJ Klasse will be Core Leader. Mr. Albert Cupo will direct the Core?s activities on a day-to-day basis. The tasks of the Core are outlined below. Other responsibilities related to the production of trimers and other relevant reagents may arise during the life of the award and will be accomplished as and when appropriate.
Aim 1 : To produce milligram quantities of high quality SOSIP trimers and related Env proteins in mammalian cell lines by transient transfection. The Core will produce, in appropriate quantities, any trimers that are designed and evaluated (in small-scale production runs) by Projects 1 and 2, and then selected for scale-up. The trimers will be assessed for purity and antigenic quality, then supplied to Projects 1 and 2, to external collaborators and to other colleagues who request our reagents.
Aim 2 : To create and use permanent cell lines expressing SOSIP trimers. The Core will use now established methods to make stable CHO, 293T or 293S cell lines expressing wild type or epitope-tagged SOSIP and germline-targeting trimers based on various genotypes, under non-GMP conditions.
Aim 3 : To guide and facilitate the production and purification of SOSIP trimers under GMP conditions, and to support clinical trials. A CMO (KBI Inc.) has successfully made multi-gram amounts of GMP-grade BG505 SOSIP.664 trimers under the direction of a consortium that also involves IAVI and the BMGF. A second translational program to produce GMP-grade GT1.1 trimers should be completed in Q2 of 2019. The Core has used its expertise to guide both programs and will continue to do so for any new ones, including by the creation and transfer of methods and the production of achievable quantities of reference standard trimers. The Core will also produce SOSIP trimers for use in clinical trial analytical programs. By providing a central service to both Research Projects, and external collaborators, the Trimer Production Core will be intimately involved in the scientific program of the entire HIVRAD team.
|Ozorowski, Gabriel; Cupo, Albert; Golabek, Michael et al. (2018) Effects of Adjuvants on HIV-1 Envelope Glycoprotein SOSIP Trimers In Vitro. J Virol 92:|
|Struwe, Weston B; Chertova, Elena; Allen, Joel D et al. (2018) Site-Specific Glycosylation of Virion-Derived HIV-1 Env Is Mimicked by a Soluble Trimeric Immunogen. Cell Rep 24:1958-1966.e5|
|Behrens, Anna-Janina; Kumar, Abhinav; Medina-Ramirez, Max et al. (2018) Integrity of Glycosylation Processing of a Glycan-Depleted Trimeric HIV-1 Immunogen Targeting Key B-Cell Lineages. J Proteome Res 17:987-999|
|Ringe, Rajesh P; Pugach, Pavel; Cottrell, Christopher A et al. (2018) Closing and opening holes in the glycan shield of HIV-1 envelope glycoprotein SOSIP trimers can redirect the neutralizing antibody response to the newly unmasked epitopes. J Virol :|
|Allen, Joel D; Sanders, Rogier W; Doores, Katie J et al. (2018) Harnessing post-translational modifications for next-generation HIV immunogens. Biochem Soc Trans 46:691-698|
|Klasse, P J; Ketas, Thomas J; Cottrell, Christopher A et al. (2018) Epitopes for neutralizing antibodies induced by HIV-1 envelope glycoprotein BG505 SOSIP trimers in rabbits and macaques. PLoS Pathog 14:e1006913|
|de Taeye, Steven W; de la Peña, Alba Torrents; Vecchione, Andrea et al. (2018) Stabilization of the gp120 V3 loop through hydrophobic interactions reduces the immunodominant V3-directed non-neutralizing response to HIV-1 envelope trimers. J Biol Chem 293:1688-1701|
|Dey, Antu K; Cupo, Albert; Ozorowski, Gabriel et al. (2018) cGMP production and analysis of BG505 SOSIP.664, an extensively glycosylated, trimeric HIV-1 envelope glycoprotein vaccine candidate. Biotechnol Bioeng 115:885-899|
|Torrents de la Peña, Alba; Sanders, Rogier W (2018) Stabilizing HIV-1 envelope glycoprotein trimers to induce neutralizing antibodies. Retrovirology 15:63|
|Medina-Ramírez, Max; Garces, Fernando; Escolano, Amelia et al. (2017) Design and crystal structure of a native-like HIV-1 envelope trimer that engages multiple broadly neutralizing antibody precursors in vivo. J Exp Med 214:2573-2590|
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