Core A (Trimer Production): Principal Investigator/Program Director: Klasse, PJ / Moore, John P. Abstract The goal of the Trimer Production Core is to support the work of the Research Projects by making appropriate quantities of high quality, cleaved, soluble envelope glycoprotein trimers and related reagents, and providing them to the Project Leaders and external collaborators. Dr. PJ Klasse will be Core Leader, with Mr. Albert Cupo directing 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 antigens may arise during the life of the HIVRAD program 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 Project 1, and then selected for scale-up. The trimers will be assessed for purity and antigenic quality, in liaison with Project 1, and supplied to Projects 1 and 2 and collaborators.
Aim 2 : To create and use permanent cell lines expressing SOSIP.664 trimers. The Core will use now established methods to make stable CHO, 293T or 293S cell lines expressing wild type or epitope-tagged SOSIP.664 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. A CMO (Catalent Inc.) will make large quantities of GMP-grade BG505 SOSIP.664 trimers under the direction of a consortium that also involves IAVI, BMGF and the Scripps CHAVI-ID. The Core will use its accumulated expertise to guide this process, including the creation and transfer of methods and the production of achievable quantities of reference standard trimers. 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.
Core A (Trimer Production): Principal Investigator/Program Director: Klasse, PJ / Moore, John P. Narrative The overall objective of this multi-site HIVRAD application is to help develop a preventative HIV-1 vaccine based on the induction of virus neutralizing antibodies by rationally designed, structurally relevant envelope glycoprotein trimers. Core A will make the SOSIP trimers designed in Projects 1 and 2, and provide them to both project teams and external collaborators. The goals of the application are highly relevant to public health and human welfare.
|Koch, Kathrin; Kalusche, Sarah; Torres, Jonathan L et al. (2017) Selection of nanobodies with broad neutralizing potential against primary HIV-1 strains using soluble subtype C gp140 envelope trimers. Sci Rep 7:8390|
|Ringe, Rajesh P; Ozorowski, Gabriel; Yasmeen, Anila et al. (2017) Improving the Expression and Purification of Soluble, Recombinant Native-Like HIV-1 Envelope Glycoprotein Trimers by Targeted Sequence Changes. J Virol 91:|
|Acharya, Kriti; Rashad, Adel A; Moraca, Francesca et al. (2017) Recognition of HIV-inactivating peptide triazoles by the recombinant soluble Env trimer, BG505 SOSIP.664. Proteins 85:843-851|
|Sanders, Rogier W; Moore, John P (2017) Native-like Env trimers as a platform for HIV-1 vaccine design. Immunol Rev 275:161-182|
|Lee, Jeong Hyun; Andrabi, Raiees; Su, Ching-Yao et al. (2017) A Broadly Neutralizing Antibody Targets the Dynamic HIV Envelope Trimer Apex via a Long, Rigidified, and Anionic ?-Hairpin Structure. Immunity 46:690-702|
|Ozorowski, Gabriel; Pallesen, Jesper; de Val, Natalia et al. (2017) Open and closed structures reveal allostery and pliability in the HIV-1 envelope spike. Nature 547:360-363|
|Behrens, Anna-Janina; Harvey, David J; Milne, Emilia et al. (2017) Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. J Virol 91:|
|Sullivan, Jonathan T; Sulli, Chidananda; Nilo, Alberto et al. (2017) High-Throughput Protein Engineering Improves the Antigenicity and Stability of Soluble HIV-1 Envelope Glycoprotein SOSIP Trimers. J Virol 91:|
|Ward, Andrew B; Wilson, Ian A (2017) The HIV-1 envelope glycoprotein structure: nailing down a moving target. Immunol Rev 275:21-32|
|van Haaren, Marlies M; van den Kerkhof, Tom L G M; van Gils, Marit J (2017) Natural infection as a blueprint for rational HIV vaccine design. Hum Vaccin Immunother 13:229-236|
Showing the most recent 10 out of 41 publications