Handlogten, Michael W.Kiziltepe, TanyelAlves, Nathan J.Bilgicer, Basar2019-03-272019-03-272012Handlogten, M. W., Kiziltepe, T., Alves, N. J., & Bilgicer, B. (2012). Synthetic allergen design reveals the significance of moderate affinity epitopes in mast cell degranulation. ACS chemical biology, 7(11), 1796-1801. http://dx.doi.org/10.1021/cb300193fhttps://hdl.handle.net/1805/18694This study describes the design of a well-defined homotetravalent synthetic allergen (HTA) system to investigate the effect of hapten–IgE interactions on mast cell degranulation. A library of DNP variants with varying affinities for IgEDNP was generated (Kd from 8.1 nM to 9.2 μM), and 8 HTAs spanning this range were synthesized via conjugation of each DNP variant to the tetravalent scaffold. HTAs with hapten Kd < 235 nM stimulated degranulation following a bell-shaped dose response curve with maximum response occurring near the hapten Kd. HTAs with hapten Kd ≥ 235 nM failed to stimulate degranulation. To mimic physiological conditions, the percent of allergen specific IgE on cell surface was varied, and maximum degranulation occurred at 25% IgEDNP. These results demonstrated that moderate hapten–IgE affinities are sufficient to trigger mast cell degranulation. Moreover, this study established the HTA design as a well-defined, controllable, and physiologically relevant experimental system to elucidate the mast cell degranulation mechanism.enhomotetravalent synthetic allergenmast cell degranulationsynthetic allergen designArticle10.1021/cb300193f