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.
BACKGROUND: Allergen-specific IgE and IgG antibodies play pivotal roles in the induction and progression of allergichypersensitivity reactions. Consequently, monoclonal humanIgE 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: M Levin; S Rotthus; S Wendel; N Najafi; E Källström; M Focke-Tejkl; R Valenta; S Flicker; M Ohlin Journal: Clin Exp Allergy Date: 2014-11 Impact factor: 5.018
Authors: Simon Blank; Stefanie Etzold; Ulf Darsow; Maximilian Schiener; Bernadette Eberlein; Dennis Russkamp; Sara Wolf; Anke Graessel; Tilo Biedermann; Markus Ollert; Carsten B Schmidt-Weber Journal: Hum Vaccin Immunother Date: 2017-05-11 Impact factor: 3.452
Authors: Frederic Jabs; Melanie Plum; Nick S Laursen; Rasmus K Jensen; Brian Mølgaard; Michaela Miehe; Marco Mandolesi; Michèle M Rauber; Wolfgang Pfützner; Thilo Jakob; Christian Möbs; Gregers R Andersen; Edzard Spillner Journal: Nat Commun Date: 2018-01-02 Impact factor: 14.919