BACKGROUND: Testing serum samples for total and allergen-specific IgE requires separate testing for each antibody and allergen specificity. OBJECTIVE: To apply fluorescent suspension array technology to allow simultaneous detection of total and allergen-specific IgE in serum in a single quantitative test. METHODS: A 7-plex suspension array for the simultaneous detection of total IgE and IgE specific to Der p 1, Der p 2, Fel d 1, Can f 1, Bet v 1, and Phl p 5 was developed, using mAb or purified allergens covalently coupled to fluorescent microspheres. The multiplex array was validated by comparing total and allergen-specific IgE levels in serum from patients with allergy with results obtained by enzyme immunoassays. RESULTS: There was a highly significant correlation between total IgE levels measured by multiplex array and fluorescent enzyme immunoassay (r = 0.97; P < .001; n = 63). Total and allergen-specific IgE levels also correlated with enzyme-linked and fluorescent enzyme immunoassay results (r = 0.44-0.94; n = 95 or 106). The multiplex array was reproducible (r = 0.86-0.99; mean coefficient of variance percentage, 12% to 25%). The sample volume required for a 7-plex assay was <20 microL per sample, compared with >400 microL in current immunoassays. CONCLUSION: The multiplex array is a high-throughput system that allows simultaneous quantification of allergen-specific and total IgE. CLINICAL IMPLICATIONS: Our results suggest that fluorescent multiplex technology will facilitate large-scale epidemiologic studies of allergic sensitization. The reduced serum volume is an advantage for pediatric studies.
BACKGROUND: Testing serum samples for total and allergen-specific IgE requires separate testing for each antibody and allergen specificity. OBJECTIVE: To apply fluorescent suspension array technology to allow simultaneous detection of total and allergen-specific IgE in serum in a single quantitative test. METHODS: A 7-plex suspension array for the simultaneous detection of total IgE and IgE specific to Der p 1, Der p 2, Fel d 1, Can f 1, Bet v 1, and Phl p 5 was developed, using mAb or purified allergens covalently coupled to fluorescent microspheres. The multiplex array was validated by comparing total and allergen-specific IgE levels in serum from patients with allergy with results obtained by enzyme immunoassays. RESULTS: There was a highly significant correlation between total IgE levels measured by multiplex array and fluorescent enzyme immunoassay (r = 0.97; P < .001; n = 63). Total and allergen-specific IgE levels also correlated with enzyme-linked and fluorescent enzyme immunoassay results (r = 0.44-0.94; n = 95 or 106). The multiplex array was reproducible (r = 0.86-0.99; mean coefficient of variance percentage, 12% to 25%). The sample volume required for a 7-plex assay was <20 microL per sample, compared with >400 microL in current immunoassays. CONCLUSION: The multiplex array is a high-throughput system that allows simultaneous quantification of allergen-specific and total IgE. CLINICAL IMPLICATIONS: Our results suggest that fluorescent multiplex technology will facilitate large-scale epidemiologic studies of allergic sensitization. The reduced serum volume is an advantage for pediatric studies.
Authors: Maksymilian Chruszcz; Anna Pomés; Jill Glesner; Lisa D Vailes; Tomasz Osinski; Przemyslaw J Porebski; Karolina A Majorek; Peter W Heymann; Thomas A E Platts-Mills; Wladek Minor; Martin D Chapman Journal: J Biol Chem Date: 2011-12-30 Impact factor: 5.157
Authors: Judith A Woodfolk; Jill Glesner; Paul W Wright; Christopher L Kepley; Mi Li; Martin Himly; Lyndsey M Muehling; Alla Gustchina; Alexander Wlodawer; Martin D Chapman; Anna Pomés Journal: J Biol Chem Date: 2015-12-07 Impact factor: 5.157
Authors: Mi Li; Alla Gustchina; Jill Glesner; Sabina Wünschmann; Lisa D Vailes; Martin D Chapman; Anna Pomés; Alexander Wlodawer Journal: J Immunol Date: 2010-12-01 Impact factor: 5.422
Authors: Ignacio J Ansotegui; Giovanni Melioli; Giorgio Walter Canonica; Luis Caraballo; Elisa Villa; Motohiro Ebisawa; Giovanni Passalacqua; Eleonora Savi; Didier Ebo; R Maximiliano Gómez; Olga Luengo Sánchez; John J Oppenheimer; Erika Jensen-Jarolim; David A Fischer; Tari Haahtela; Martti Antila; Jean J Bousquet; Victoria Cardona; Wen Chin Chiang; Pascal M Demoly; Lawrence M DuBuske; Marta Ferrer Puga; Roy Gerth van Wijk; Sandra Nora González Díaz; Alexei Gonzalez-Estrada; Edgardo Jares; Ayse Füsun Kalpaklioğlu; Luciana Kase Tanno; Marek L Kowalski; Dennis K Ledford; Olga Patricia Monge Ortega; Mário Morais Almeida; Oliver Pfaar; Lars K Poulsen; Ruby Pawankar; Harald E Renz; Antonino G Romano; Nelson A Rosário Filho; Lanny Rosenwasser; Mario A Sánchez Borges; Enrico Scala; Gian-Enrico Senna; Juan Carlos Sisul; Mimi L K Tang; Bernard Yu-Hor Thong; Rudolf Valenta; Robert A Wood; Torsten Zuberbier Journal: World Allergy Organ J Date: 2020-02-25 Impact factor: 4.084