title: Conformational evaluation of HIV-1 trimeric envelope glycoproteins using a cell-based ELISA assay.
abstract: HIV-1 envelope glycoproteins (Env) mediate viral entry into target cells and are essential to the infectious cycle. Understanding how those glycoproteins are able to fuel the fusion process through their conformational changes could lead to the design of better, more effective immunogens for vaccine strategies. Here we describe a cell-based ELISA assay that allows studying the recognition of trimeric HIV-1 Env by monoclonal antibodies. Following expression of HIV-1 trimeric Env at the surface of transfected cells, conformation specific anti-Env antibodies are incubated with the cells. A horseradish peroxidase-conjugated secondary antibody and a simple chemiluminescence reaction are then used to detect bound antibodies. This system is highly flexible and can detect Env conformational changes induced by soluble CD4 or cellular proteins. It requires minimal amount of material and no highly-specialized equipment or know-how. Thus, this technique can be established for medium to high throughput screening of antigens and antibodies, such as newly-isolated antibodies.
authors.affiliation: Centre de recherche du CHUM, Department of Microbiology, Infectiology and Immunology, Université de Montréal.
authors.forename: Maxime
authors.initials: M
authors.lastname: Veillette
authors.affiliation: Centre de recherche du CHUM, Department of Microbiology, Infectiology and Immunology, Université de Montréal.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Coutu
authors.affiliation: Centre de recherche du CHUM, Department of Microbiology, Infectiology and Immunology, Université de Montréal.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Richard
authors.affiliation: Centre de recherche du CHUM, Department of Microbiology, Infectiology and Immunology, Université de Montréal.
authors.forename: Laurie-Anne
authors.initials: LA
authors.lastname: Batraville
authors.affiliation: Centre de recherche du CHUM, Department of Microbiology, Infectiology and Immunology, Université de Montréal.
authors.forename: Anik
authors.initials: A
authors.lastname: Désormeaux
authors.affiliation: Centre de recherche du CHUM, Department of Microbiology, Infectiology and Immunology, Université de Montréal.
authors.forename: Michel
authors.initials: M
authors.lastname: Roger
authors.affiliation: Centre de recherche du CHUM, Department of Microbiology, Infectiology and Immunology, Université de Montréal; andres.finzi@umontreal.ca.
authors.forename: Andrés
authors.initials: A
authors.lastname: Finzi
mesh_terms: D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology; D001859:Bone Neoplasms; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015497:HIV-1 / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods; D012516:Osteosarcoma; D011487:Protein Conformation; D014162:Transfection; D054299:env Gene Products, Human Immunodeficiency Virus / Q000737:chemistry* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D054299:env Gene Products, Human Immunodeficiency Virus