title: Binding interactions between soluble HIV envelope glycoproteins and quaternary-structure-specific monoclonal antibodies PG9 and PG16.
abstract: PG9 and PG16 are antibodies isolated from a subject infected with HIV-1 and display broad anti-HIV neutralizing activities. They recognize overlapping epitopes, which are preferentially expressed on the membrane-anchored trimeric form of the HIV envelope glycoprotein (Env). PG9 and PG16 were reported not to bind to soluble mimetics of Env. The engineering of soluble Env proteins on which the PG9 and PG16 epitopes are optimally exposed will support efforts to elicit broad anti-HIV neutralizing antibodies by immunization. Here, we identified several soluble gp140 Env proteins that are recognized by PG9 and PG16, and we investigated the molecular details of those binding interactions. The IgG versions of PG9 and PG16 recognize the soluble trimeric gp140 form less efficiently than the corresponding monomeric gp140 form. In contrast, the Fab versions of PG9 and PG16 recognized the monomeric and trimeric gp140 forms with identical binding kinetics and with binding affinities similar to the high binding affinity of the anti-V3 antibody 447D to its epitope. Our data also indicate that, depending on the Env backbone, the interactions of PG9 and PG16 with gp140 may be facilitated by the presence of the gp41 ectodomain and are independent of the proper enzymatic cleavage of gp140 into gp120 and gp41. The identification of soluble Env proteins that express the PG9 and PG16 epitopes and the detailed characterization of the molecular interactions between these two antibodies and their ligands provide important and novel information that will assist in improving the engineering of future Env immunogens.
authors.affiliation: Seattle Biomedical Research Institute, 307 Westlake Avenue North, Seattle, WA 98109, USA.
authors.forename: Thaddeus M
authors.initials: TM
authors.lastname: Davenport
authors.forename: Della
authors.initials: D
authors.lastname: Friend
authors.forename: Katharine
authors.initials: K
authors.lastname: Ellingson
authors.forename: Hengyu
authors.initials: H
authors.lastname: Xu
authors.forename: Zachary
authors.initials: Z
authors.lastname: Caldwell
authors.forename: George
authors.initials: G
authors.lastname: Sellhorn
authors.forename: Zane
authors.initials: Z
authors.lastname: Kraft
authors.forename: Roland K
authors.initials: RK
authors.lastname: Strong
authors.forename: Leonidas
authors.initials: L
authors.lastname: Stamatatos
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D006801:Humans; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D012995:Solubility; D020349:Surface Plasmon Resonance; D054299:env Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; D054299:env Gene Products, Human Immunodeficiency Virus; C097729:gp140 envelope protein, Human immunodeficiency virus 1