title: Rapid production of antigen-specific monoclonal antibodies from a variety of animals.
abstract: BACKGROUND
  Although a variety of animals have been used to produce polyclonal antibodies against antigens, the production of antigen-specific monoclonal antibodies from animals remains challenging.
  
  
  RESULTS
  We propose a simple and rapid strategy to produce monoclonal antibodies from a variety of animals. By staining lymph node cells with an antibody against immunoglobulin and a fluorescent dye specific for the endoplasmic reticulum, plasma/plasmablast cells were identified without using a series of antibodies against lineage markers. By using a fluorescently labeled antigen as a tag for a complementary cell surface immunoglobulin, antigen-specific plasma/plasmablast cells were sorted from the rest of the cell population by fluorescence-activated cell sorting. Amplification of cognate pairs of immunoglobulin heavy and light chain genes followed by DNA transfection into 293FT cells resulted in the highly efficient production of antigen-specific monoclonal antibodies from a variety of immunized animals.
  
  
  CONCLUSIONS
  Our technology eliminates the need for both cell propagation and screening processes, offering a significant advantage over hybridoma and display strategies.
authors.affiliation: Laboratory of Molecular and Cellular Biology, Faculty of Science and Engineering, Graduate School, University of Toyama, 3190 Gofuku, Toyama-shi, Toyama, 930-8555, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Kurosawa
authors.forename: Megumi
authors.initials: M
authors.lastname: Yoshioka
authors.forename: Rika
authors.initials: R
authors.lastname: Fujimoto
authors.forename: Fuminori
authors.initials: F
authors.lastname: Yamagishi
authors.forename: Masaharu
authors.initials: M
authors.lastname: Isobe
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity / Q000276:immunology*; D015415:Biomarkers / Q000378:metabolism; D019070:Cell Lineage / Q000276:immunology; D002469:Cell Separation; D004721:Endoplasmic Reticulum / Q000378:metabolism; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology; D007328:Insulin / Q000276:immunology; D010802:Phylogeny; D010950:Plasma Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D000939:Epitopes; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D007328:Insulin