title: The structure of dual-variable-domain immunoglobulin molecules alone and bound to antigen.
abstract: A dual-specific, tetravalent immunoglobulin G-like molecule, termed dual variable domain immunoglobulin (DVD-Ig™), is engineered to block two targets. Flexibility modulates Fc receptor and complement binding, but could result in undesirable cross-linking of surface antigens and downstream signaling. Understanding the flexibility of parental mAbs is important for designing and retaining functionality of DVD-Ig™ molecules. The architecture and dynamics of a DVD-Ig™ molecule and its parental mAbs was examined using single particle electron microscopy. Hinge angles measured for the DVD-Ig™ molecule were similar to the inner antigen parental mAb. The outer binding domain of the DVD-Ig™ molecule was highly mobile and three-dimensional (3D) analysis showed binding of inner antigen caused the outer domain to fold out of the plane with a major morphological change. Docking high-resolution X-ray structures into the 3D electron microscopy map supports the extraordinary domain flexibility observed in the DVD-Ig™ molecule allowing antigen binding with minimal steric hindrance.
authors.affiliation: Protein Analytics; AbbVie Bioresearch Center; Worcester, MA USA.
authors.forename: Ivan
authors.initials: I
authors.lastname: Correia
authors.affiliation: NanoImaging Services, Inc.; La Jolla, CA USA.
authors.forename: Joyce
authors.initials: J
authors.lastname: Sung
authors.affiliation: Protein Analytics; AbbVie Bioresearch Center; Worcester, MA USA.
authors.forename: Randall
authors.initials: R
authors.lastname: Burton
authors.affiliation: Department of Structural Biology; AbbVie Laboratories; Abbott Park, IL USA.
authors.forename: Clarissa G
authors.initials: CG
authors.lastname: Jakob
authors.affiliation: NanoImaging Services, Inc.; La Jolla, CA USA.
authors.forename: Bridget
authors.initials: B
authors.lastname: Carragher
authors.affiliation: Biologics; AbbVie Bioresearch Center; Worcester, MA USA.
authors.forename: Tariq
authors.initials: T
authors.lastname: Ghayur
authors.affiliation: Protein Analytics; AbbVie Bioresearch Center; Worcester, MA USA.
authors.forename: Czeslaw
authors.initials: C
authors.lastname: Radziejewski
mesh_terms: D018033:Antibodies, Bispecific / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000627:therapeutic use; D000941:Antigens / Q000276:immunology; D018360:Crystallography, X-Ray; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry*; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D007167:Immunotherapy; D018664:Interleukin-12 / Q000737:chemistry / Q000276:immunology; D020382:Interleukin-18 / Q000737:chemistry / Q000276:immunology; D046529:Microscopy, Electron, Transmission; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods; D017434:Protein Structure, Tertiary
chemical_list: D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D000941:Antigens; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D020382:Interleukin-18; D018664:Interleukin-12
keywords: bispecific antibodies; crystallography; dual variable domain molecules; immunoglobulin; structure; transmission electron microscopy