title: Generation of human monoclonal allergen-specific IgE and IgG antibodies from synthetic antibody libraries.
abstract: BACKGROUND
  Allergen-specific IgE and IgG antibodies play pivotal roles in the induction and progression of allergic hypersensitivity reactions. Consequently, monoclonal human IgE and IgG4 antibodies with defined specificity for allergens should be useful in allergy research and diagnostic tests. We used combinatorial antibody libraries and subsequent recombinant production to make and assess IgE, IgG1, and IgG4 allergen-specific antibodies.
  
  
  METHODS
  We used phage display to select a synthetic single-chain antibody fragment (scFv) library against 3 different allergens, from bee venom, bovine milk, and apple. The scFv obtained were converted into IgG1, IgG4, and IgE antibody formats and assessed for their biochemical properties by ELISA, immunoblotting, and fluorescence-activated cell sorting.
  
  
  RESULTS
  Two different antibody formats for each IgG1, IgG4, and IgE antibody were produced in mammalian cells as disulfide-linked and glycosylated Ig, which were usable in allergen-specific ELISA assays and immunoblots. In addition, the recombinant IgE antibodies mediated the binding of allergens to HEK-293 cells transfected with the high-affinity IgE receptor, and this binding was blocked by corresponding IgG antibodies.
  
  
  CONCLUSIONS
  The use of synthetic libraries for the generation of allergen-specific recombinant IgE and IgG antibodies should have broad applications in allergological research and diagnosis.
authors: Institute of Biochemistry and Food Sciences, Division of Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany.
authors: Ingke
authors: I
authors: Braren
authors: Simon
authors: S
authors: Blank
authors: Henning
authors: H
authors: Seismann
authors: Susanne
authors: S
authors: Deckers
authors: Markus
authors: M
authors: Ollert
authors: Thomas
authors: T
authors: Grunwald
authors: Edzard
authors: E
authors: Spillner
mesh_terms: D000485:Allergens / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D000918:Antibody Specificity; D001514:Bee Venoms / Q000276:immunology; D002417:Cattle; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D027845:Malus / Q000276:immunology; D008892:Milk / Q000276:immunology; D019151:Peptide Library; D021122:Protein Subunits / Q000378:metabolism; D017455:Receptors, IgE / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D000485:Allergens; D000911:Antibodies, Monoclonal; D001514:Bee Venoms; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D021122:Protein Subunits; D017455:Receptors, IgE; D011994:Recombinant Proteins; D007073:Immunoglobulin E