Generation of Human Monoclonal Allergen-Specific IgE and IgG Antibodies from Synthetic Antibody Libraries

@article{citeulike:2625456, 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. 10.1373/clinchem.2006.078360}, author = {Braren, Ingke and Blank, Simon and Seismann, Henning and Deckers, Susanne and Ollert, Markus and Grunwald, Thomas and Spillner, Edzard }, citeulike-article-id = {2625456}, doi = {10.1373/clinchem.2006.078360}, journal = {Clin Chem}, keywords = {methods}, month = {May}, number = {5}, pages = {837--844}, posted-at = {2008-04-03 09:37:54}, priority = {2}, title = {Generation of Human Monoclonal Allergen-Specific IgE and IgG Antibodies from Synthetic Antibody Libraries}, url = {http://dx.doi.org/10.1373/clinchem.2006.078360}, volume = {53}, year = {2007} }

TY - JOUR ID - citeulike:2625456 L3 - citeulike-article-id:2625456 TI - Generation of Human Monoclonal Allergen-Specific IgE and IgG Antibodies from Synthetic Antibody Libraries JF - Clin Chem VL - 53 IS - 5 SP - 837 EP - 844 N2 - 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. 10.1373/clinchem.2006.078360 KW - methods AU - Braren, Ingke AU - Blank, Simon AU - Seismann, Henning AU - Deckers, Susanne AU - Ollert, Markus AU - Grunwald, Thomas AU - Spillner, Edzard PY - 2007/05/01/ UR - http://dx.doi.org/10.1373/clinchem.2006.078360 ER -