title: Purification and identification of cell surface antigens using lamprey monoclonal antibodies.
abstract: Variable lymphocyte receptor (VLR) B antibodies of the evolutionary distant sea lamprey are structurally distinct from conventional mammalian antibodies. The different protein architecture and large evolutionary distance of jawless vertebrates suggest that VLR antibodies may represent promising tools for biomarker discovery. Here we report the generation of panels of monoclonal VLR antibodies from lamprey larvae immunized with human T cells and the use of a recombinant monoclonal VLR antibody for antigen purification and mass spectrometric identification. We demonstrate that despite predicted low affinity of individual VLR antigen binding units to the antigen, the high avidity resulting from decameric assembly of secreted VLR antibodies allows for efficient antigen capture and subsequent identification by mass spectometry. We show that VLR antibodies detect their antigens with high specificity and can be used in various standard laboratory application techniques. The lamprey antibodies are novel reagents that can complement conventional monoclonal antibodies in multiple scientific research disciplines.
authors.affiliation: Department of Pathology and Laboratory Medicine, Emory University School of Medicine, 1462 Clifton Rd NE, Room 403C, Atlanta, GA 30322, USA.
authors.forename: Cuiling
authors.initials: C
authors.lastname: Yu
authors.forename: Shabab
authors.initials: S
authors.lastname: Ali
authors.forename: Jonathan
authors.initials: J
authors.lastname: St-Germain
authors.forename: Yanling
authors.initials: Y
authors.lastname: Liu
authors.forename: Xuecong
authors.initials: X
authors.lastname: Yu
authors.forename: David L
authors.initials: DL
authors.lastname: Jaye
authors.forename: Michael F
authors.initials: MF
authors.lastname: Moran
authors.forename: Max D
authors.initials: MD
authors.lastname: Cooper
authors.forename: Götz R A
authors.initials: GR
authors.lastname: Ehrhardt
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000915:Antibody Affinity; D000954:Antigens, Surface / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007114:Immunization; D007163:Immunosorbent Techniques; D007798:Lampreys / Q000276:immunology*; D007814:Larva; D013058:Mass Spectrometry; D011485:Protein Binding; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface