Novel murine mAbs define specific and cross-reactive epitopes on the latex profilin panallergen Hev b 8
Por:
Mares-Mejía I., García-Ramírez B., Torres-Larios A., Rodríguez-Hernández A., Osornio-Hernández A.I., Terán-Olvera G., Ortega E., Rodríguez-Romero A.
Publicada:
1 ene 2020
Resumen:
The production of specific antibodies able to recognize allergens from different sources or block interactions between allergens and antibodies mediating allergic reactions is crucial for developing successful tools for diagnostics and therapeutics. Panallergens are highly conserved proteins present in widely different species, implicated in relevant cross-reactions. The panallergen latex profilin (Hev b 8) has been associated with the latex-food-pollen syndrome. We generated five monoclonal IgGs and one IgE from murine hybridomas against recombinant Hev b 8 and evaluated their interaction with this allergen using ELISA and biolayer interferometry (BLI). Affinity purified mAbs exhibited high binding affinities towards rHev b 8, with KD1 values ranging from 10-10 M to 10-11 M. Some of these antibodies also recognized the recombinant profilins from maize and tomato (Zea m 12 and Sola l 1), and the ash tree pollen (Fra e 2). Competition ELISA demonstrated that some mAb pairs could bind simultaneously to rHev b 8. Using BLI, we detected competitive, non-competitive, and partial-competition interactions between pairs of mAbs with rHev b 8, suggesting the existence of at least two non-overlapping epitopes on the surface of this allergen. Three-dimensional models of the Fv of 1B4 and 2D10 IgGs and docking simulations of these Fvs with rHev b 8 revealed these epitopes. Furthermore, these two mAbs inhibited the interaction of polyclonal IgE and IgG4 antibodies from profilin-allergic patients with rHev b 8, indicating that the mAbs and the antibodies present in sera from allergic patients bind to overlapping epitopes on the allergen. These mAbs can be useful tools for immune-localization studies, immunoassay development, or standardization of allergenic products. © 2020 Elsevier Ltd
Filiaciones:
Mares-Mejía I.:
Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
García-Ramírez B.:
Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
Torres-Larios A.:
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
Rodríguez-Hernández A.:
Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
Osornio-Hernández A.I.:
Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
Terán-Olvera G.:
Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
Ortega E.:
Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
Rodríguez-Romero A.:
Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Cd. Universitaria, Coyoacán, Ciudad de México, 04510, Mexico
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