The scarcity of monoclonal human @entity9416 antibodies with specificity for defined allergens is a bottleneck for the molecular characterisation of allergens and their epitopes. Insights into the characteristics of such antibodies may allow for analyses of the molecular basis underlying allergenicity and cross-reactivity, standardisation of allergens as well as improvement of @entity566 diagnostics and therapeutics. Here we report the generation and application of the first set of authentic human IgG, @entity9416 and IgA antibodies. On the basis of a @entity25128 p 5a specific antibody fragment, a lambda light chain and the IgG1, IgG4, @entity9416 , @entity5727 , and IgA2 heavy chains, the corresponding @entity1 immunoglobulins were constructed and produced in @entity1 cells. In parallel, a @entity19 hybridoma line with specificity for @entity25128 p 5a was established, recloned and produced as @entity1 chimeric @entity9416 . After purification, immunoreactivity of the antibodies with the allergen was assessed. Applicability in @entity566 diagnostics was confirmed by establishment of artificial @entity1 sera. Functionality of both antibodies was further demonstrated in receptor binding studies and mediator release assays using humanised @entity35 basophil @entity1778 cells (RBL-SX38) suggesting the presence of spatially separate epitopes. By using @entity25128 p 5 fusion proteins and recombinant @entity9416 in immunoblotting and mediator release assays we assigned the epitope of the authentic @entity9416 to a looped stretch exclusively present in @entity25128 p 5a. In summary, the @entity25128 p 5-specific antibodies are the first full set of @entity566 -related antibody isotypes of their kind and represent valuable tools for studies of fundamental mechanisms and structure/function relationships in @entity566 .

Generation and epitope analysis of human monoclonal antibody isotypes with specificity for the Timothy grass major allergen XXXX p 5a.

@entity5727 :: ('3493', 'Gene') :: ['IgA1'] @entity1 :: ('9606', 'Species') :: ['mammalian', 'human'] @entity19 :: ('10090', 'Species') :: ['murine'] @entity566 :: ('MESH:D006967', 'Disease') :: ['allergy'] @entity25128 :: ('613', 'Gene') :: ['Phl'] @entity1778 :: ('MESH:D007938', 'Disease') :: ['leukaemia'] @entity35 :: ('10116', 'Species') :: ['rat'] @entity9416 :: ('3497', 'Gene') :: ['IgE']
@entity25128 :: (613,Gene) :: ['Phl']
