Validación interlaboratorio del método de SDS-PAGE para la determinación de la composición de proteínas en extractos alergénicos de ácaros

La Monografía de la Farmacopea Europea para Productos Alergénicos establece la importancia de la determinación de la composición de proteínas a productos finales y también a la Materia Prima Alergénica (MPA) y el Ingrediente Farmacéutico Activo (IFA). La validación de métodos analíticos asegura la calidad del producto desde sus estadios iniciales. El objetivo de este trabajo fue validar el método de SDS-PAGE para extractos alergénicos de ácaros Dermatophagoides siboney (Ds) y Blomia tropicalis (Bt) en las tres etapas del proceso (MPA, IFA y producto final). Participaron en el estudio tres laboratorios de BIOCEN. Se evaluó la Precisión (Repetibilidad, Precisión Intermedia y Reproducibilidad) en la determinación del contenido relativo de las bandas principales que contienen componentes alergénicos. Como parámetro de Especificidad se analizó la posible interferencia del bicarbonato de amonio. Se evaluó la Exactitud y se determinó el Límite de Detección (LD) en la determinación del peso molecular de las bandas principales. Se cumplieron todos los criterios de aceptación establecidos. El método fue preciso, todos los coeficientes de variación (CV) calculados fueron 0.05. No hubo diferencias significativas entre porcentajes de las bandas principales en los extractos alergénicos con Bicarbonato de Amonio en su composición original según ANOVA, p>0.05. El método fue exacto, no hubo diferencias significativas (Prueba t Student, p>0.05) entre los valores experimentales del peso molecular y los reportados en la literatura. El LD la técnica fue de para MPA Ds 5000UB, Bt 50000UB; IFA y Producto final de Ds 2500UB, Bt 12500UB. En conclusión, quedó validada la técnica analítica SDS-PAGE para la determinación de la composición de proteínas en los extractos alergénicos de ácaros en tres etapas diferentes del proceso de fabricación. Se fortalece así el aseguramiento de la calidad de estos productos

Is peanut causing food allergy in Cuba? Preliminary assessment of allergic sensitization and IgE specificity profile to peanut allergens in Cuban allergic patients

Background Peanut allergy is increasing at an alarming pace in developed countries. Peanut (Arachis hypogaea) is a common food in Cuba. Nevertheless, reported values of sensitization and symptom severity are usually low. As our objective, we carried out an evaluation of allergic sensitivity to perform an assessment of allergic sensitization and IgE specificity profile to peanut allergens in Cuban allergic patients.Methods The Skin Prick Test (SPT) was performed for each patient, using two glycerinated allergenic extracts, prepared from raw or roasted peanuts. Overall, 316 food allergic patients (159 adults and 157 children) attending allergy services at four hospitals in Havana were included, as well as 303 adult non- allergic volunteers. The IgE binding profile of 26 selected SPT positive patients was further analyzed by immunoblotting.Results The prevalence of sensitization to peanut was 4.6% in general adult population, whereas in adult food-allergic patients it was 18.6%. Prevalence rates were even greater in food allergic children achieving 25.8%. Sensitization frequencies were apparently greater for roasted, as compared to raw peanuts, although the difference was not significant (p> 0.05, Mc Nemar's). IgE binding was shown mostly by the 15 and 17 kDa bands, tentatively identified as the major allergens Ara h 2 and Ara h 6. The IgG4 binding profile was similar to IgE, although with more prominence of the bands at 37 and 28 KDa, corresponding to an Ara h 3 fragment and Peanut Agglutinin.Discussion The study estimated a relatively high prevalence of peanut sensitization in population. Data reported here suggest that IgE sensitization in Cuban patients is focused mostly on MW bands corresponding to the major allergens Ara h 6 and Ara h 2.Conclusions Sensitization to peanut allergen is indeed relatively frequent in Cuba. The IgE profile is congruent to a sensitization pattern by ingestion of roasted peanuts and is directed to well-known major allergens. Keywords: Allergic sensitization, Peanut allergens, Food allergy, IgE specificit

Proceedings of the 2015 WAO Symposium on Food Allergy and the Microbiome

Table of contents A1 Characterization of the immunoallergic profile towards the proteins of the wheat flour in Cuban population Raúl Lázaro Castro Almarales, Mary Carmen Reyes Zamora, Beatriz Tamargo, Damaris Torralba Averoff, Raysa Cruz, Yunia Oliva Diaz, Mirta Alvarez Castello, Alexander Ciria, Alexis Labrada, Maytee Mateo A2 Are peanuts causing food allergy in Cuba? Maytee Mateo, Damaris Torralba Averoff, Raysa Cruz, Yunia Oliva Diaz, Mirta Alvarez Castello, Alexander Ciria, Mary Carmen Reyes Zamora, Beatriz Tamargo, Alexis Labrada A3 Prick test and immunoallergic profile to soy allergens in Cuban population Omar Herrera, Maytee Mateo, Raysa Cruz, Mirta Alvarez Castello, Alexander Ciria, Raúl Lázaro Castro Almarales, Mary Carmen Reyes Zamora, Alexis Labrada A4 Skin sensitization and immunoallergic profile to hen's egg in Cuban population José Severino Rodríguez Canosa, Raysa Cruz, Maytee Mateo, Mirta Alvarez Castello, Alexander Ciria, Raúl Lázaro Castro Almarales, Mary Carmen Reyes Zamora, Alexis Labrada A5 Sensitization to three domestic mites in patients with adverse food events to shellfish Mirta Alvarez Castello, Raúl Lázaro Castro Almarales, Alexis Labrada, Biocen A6 Diagnostic efficacy by skin prick test with allergenic extracts of legumes in Cuban patients Yamilet Ibizate Novales, Ilonka Estruch Fajardo, Alexis Labrada, Maytee Mateo, Armando Ginard A7 Baked egg goods without wheat flour carry an increased risk of reaction Bruce Lanser, Anna Faino, Erwin Gelfand, Pia Hauk A8 Prevalence, incidence and associated risk factors of adverse reaction to food in Cuban infants - a population-based prospective study Silvia Venero Fernández, Julia Urbina, Mirta Alvarez Castello, Raúl Lázaro Castro Almarales, Ramón Suárez Medina, Hermes Fundora Hernández, John Britton, Andrew William Fogarty A9 Microbiome in ice machines and assessing the plasma nanotechnology in breaking the biofilm and improving air quality Nabarun Ghosh, Clinton Ross Bell, Chandini Revanna, Constantine Saadeh, Jeff Bennert, Danius Bouyi, Mitsy Veloz, Nelofar Sherali A10 Characteristics of patients with food allergy in health public service Magna Coelho A11 Allergic rhinitis and asthma index increased in Texas panhandle and AHPCO and plasma nanotechnology as solutions Nabarun Ghosh, Jeff Bennert, Danius Bouyi, Constantine Saadeh, Clinton Ross Bell, Mitsy Veloz, Chandini Revanna, Nelofar Sherali A12 Antigen-specific T follicular helper cells mediate peanut allergy in mice Joseph J. Dolence, Takao Kobayashi, Koji Iijima, Hirohito Kita, Hirohito Kita, Ashli Moore, James Krempski A13 Production of recombinant Mal d 3, a major apple allergen, in Pichia Pastoris, to investigate the impact of the food matrix and post-translational modifications on Mal d 3 immuno-reactivity Roberta Aina, Riccardo Asero, Sabine Pfeifer, Pawel Dubiela, Merima Bublin, Christian Radauer, Piotr Humeniuk, Karin Hoffmann-Sommergruber A14 Reaction to sports drink: no whey! Whey allergy in absence of clinical cow’s milk allergy Frank Eidelman, Ves Dimov, Charl Khalil A15 Food allergy on Tumblr: focus on teenage audience may increase educational impact Ves Dimov, Frank Eidelman, Charl Khalil A16 Changes in IgE levels following one-year immunizations in two children with food allergy Alice E. W. Hoyt, Peter Heymann, Alexander Schuyler, Scott Commins, Thomas Platts-Mills A17 IgE and IgG4 antibodies to cow's milk components in children with eosinophilic esophagitis: higher specific IgG4 antibodies and IgG4:IgE ratios compared with subjects with IgE-mediated food allergy Alexander Schuyler, Patrice Kruszewski, John Russo, Lisa Workman, Thomas Platts-Mills, Elizabeth Erwin, Anubha Tripathi A18 Frequency of Sensitization to Food Allergens in Patients with Rhinitis and Asthma in the National Medical Center La Raza “Dr. Antonio Fraga Mouret”, Mexico City Gabriela Yvette Castellanos, Elizabeth Mendieta, Martín Becerril-Angele