title: Construction and characterization of soluble, cleaved, and stabilized trimeric Env proteins based on HIV type 1 Env subtype A.
abstract: The generation of an antibody response capable of neutralizing a broad range of clinical isolates remains an important goal of human immunodeficiency virus type 1 (HIV-1) vaccine development. Envelope glycoprotein (Env)-based vaccine candidates will also need to take into account the extensive genetic diversity of circulating HIV-1 strains. We describe here the generation of soluble, stabilized, proteolytically cleaved, trimeric forms of Env (SOSIP gp140 proteins) based on contemporary Env subtype A viruses from East Africa. We discuss issues associated with the construction, purification, and characterization of such complex proteins; not all env sequences allow the expression of trimeric proteins. However, stabilized trimers from one such protein, KNH1144 SOSIP gp140, were successfully made. These proteins are now being prepared for preclinical immunogenicity studies.
authors.affiliation: Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, New York 10021, USA.
authors.forename: Simon
authors.initials: S
authors.lastname: Beddows
authors.forename: Marc
authors.initials: M
authors.lastname: Kirschner
authors.forename: Lila
authors.initials: L
authors.lastname: Campbell-Gardener
authors.forename: Michael
authors.initials: M
authors.lastname: Franti
authors.forename: Antu K
authors.initials: AK
authors.lastname: Dey
authors.forename: Sai Prasad N
authors.initials: SP
authors.lastname: Iyer
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Maddon
authors.forename: Maciej
authors.initials: M
authors.lastname: Paluch
authors.forename: Aditi
authors.initials: A
authors.lastname: Master
authors.forename: Julie
authors.initials: J
authors.lastname: Overbaugh
authors.forename: Thomas
authors.initials: T
authors.lastname: VanCott
authors.forename: William C
authors.initials: WC
authors.lastname: Olson
authors.forename: John P
authors.initials: JP
authors.lastname: Moore
mesh_terms: D016915:AIDS Vaccines; D000351:Africa, Eastern; D000818:Animals; D002460:Cell Line; D019281:Dimerization; D015195:Drug Design; D015686:Gene Products, env / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D015483:HIV Antibodies / Q000097:blood*; D015497:HIV-1 / Q000145:classification; D006801:Humans; D051379:Mice; D009500:Neutralization Tests; D011817:Rabbits; D054299:env Gene Products, Human Immunodeficiency Virus
chemical_list: D016915:AIDS Vaccines; D015686:Gene Products, env; D015483:HIV Antibodies; D054299:env Gene Products, Human Immunodeficiency Virus; C097729:gp140 envelope protein, Human immunodeficiency virus 1