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A novel bioactivity assay for monoclonal antibodies directed against IgE

1994; Elsevier BV; Volume: 171; Issue: 2 Linguagem: Inglês

10.1016/0022-1759(94)90039-6

1872-7905

David T.W. Fei, John R. Lowe, Paula Jardieu,

Food Allergy and Anaphylaxis Research

A novel bioactivity assay has been developed to quantitate the biological activity of a humanized, monoclonal anti-IgE antibody (rhuMAbE25) in human whole blood. Heparinized blood specimens from prescreened healthy donors were sensitized for 2 h with a constant amount of human plasma containing IgE specific for ragweed and then challenged with ragweed allergen. Histamine was released in a dose-dependent fashion and reached plateau levels after 30 min. As expected, the release of histamine by ragweed allergen was time, temperature and Ca2+ dependent, and could be enhanced by the presence of 33% deuterium oxide. Allergen-triggered release could be inhibited by rhuMAbE25 with an effective dose range from 0.1 to 1 microgram/ml. Preincubation with other humanized MAbs, which exhibit 95% homology to rhuMAbE25 but differ in epitope specificity, failed to inhibit the ragweed-induced histamine release. Overall, this bioactivity assay has a low interassay variability (%CV) of 17% (n = 23) and can be readily modified to determine if rhuMAbE25 or other anti-allergy therapeutics are capable of blocking histamine release elicited by other allergens. Moreover, the assay can be used to confirm IgE-mediated allergic responses and to provide early information regarding safety and potential efficacy of therapeutics aimed at blocking IgE dependent responses.