Intrauterine sensitization of allergen-specific IgE analyzed by a highly sensitive new allergen microarray Norio Kamemura, MSc, Hitomi Tada, MSc, Naoki.

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Intrauterine sensitization of allergen-specific IgE analyzed by a highly sensitive new allergen microarray Norio Kamemura, MSc, Hitomi Tada, MSc, Naoki Shimojo, MD, PhD, Yoshinori Morita, MD, PhD, Yoichi Kohno, MD, PhD, Takao Ichioka, MD, Koichi Suzuki, MSc, Kenji Kubota, MSc, Mineyoshi Hiyoshi, PhD, Hiroshi Kido, MD, PhD Journal of Allergy and Clinical Immunology Volume 130, Issue 1, Pages 113-121.e2 (July 2012) DOI: 10.1016/j.jaci.2012.02.023 Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Multiallergen-specific profiling patterns of IgE, IgA, IgG, and IgG4 in paired CB, NB, and MB analyzed by the DLC chip. A, Rainbow displays of fluorescence intensities of allergen-specific IgE, IgA, IgG, and IgG4 reactivities against allergens of an array probed sera of paired CB, NB, and MB. B, Layout of the allergen DLC chip. Df, Dermatophagoides farinae; Dp, Dermatophagoides pteronyssinus; GD, guideline dot by standard serum; Std, standard calibration of human serum IgE from the National Institute for Biological Standards and Control. Each allergen was spotted in quadruple on the chip. Results are representative of 1 pair of 92 samples. Journal of Allergy and Clinical Immunology 2012 130, 113-121.e2DOI: (10.1016/j.jaci.2012.02.023) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Allergen-specific IgE adsorption for evaluating cross-reactivity of antigen-IgE antibody reaction on the DLC chip. MB sample containing allergen-specific IgE against both food and inhalant allergens was preincubated with egg white (5 μg/mL), milk (5 μg/mL), Df (500 μg/mL), or cedar pollen (500 μg/mL) at 37°C for 2 hours. After the reaction, each sample was centrifuged at 17,500 × g for 30 minutes to remove antigen-antibody complex, and then the supernatant was used for multiallergen profiling of IgE on the DLC chip. Bottom, layout of the allergen on the DLC chip. Df, Dermatophagoides farinae; Dp, Dermatophagoides pteronyssinus; GD, guideline dot; Std, standard calibration of human serum IgE. Journal of Allergy and Clinical Immunology 2012 130, 113-121.e2DOI: (10.1016/j.jaci.2012.02.023) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Total IgA levels in CB and MB. Total IgA levels in CB and MB (n = 92) were analyzed by ELISA. Journal of Allergy and Clinical Immunology 2012 130, 113-121.e2DOI: (10.1016/j.jaci.2012.02.023) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Comparison and correlation of allergen-specific IgE levels between CB and MB analyzed by the DLC chip. Allergen-specific IgE levels in CB and MB (n = 92) depicted in BUe. The cutoff value was 10 BUe. A, Milk. B, α-Casein. C, Ovomucoid. D, Cedar pollen. E, Dp. F, Df. Spearman's rank correlation test was used to assess the relation between the values of CB and MB. Df, Dermatophagoides farinae; Dp, Dermatophagoides pteronyssinus. Journal of Allergy and Clinical Immunology 2012 130, 113-121.e2DOI: (10.1016/j.jaci.2012.02.023) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Comparison of allergen-specific IgG levels in CB and MB analyzed by the DLC chip. Allergen-specific IgG levels in paired samples of CB and MB (n = 92) depicted in BUg. The cutoff value was 2.50 BUg. A, Milk. B, α-Casein. C, Ovomucoid. D, Dp. E, Df. Spearman's rank correlation test was used to assess the relation between CB and MB. Df, Dermatophagoides farinae; Dp, Dermatophagoides pteronyssinus. Journal of Allergy and Clinical Immunology 2012 130, 113-121.e2DOI: (10.1016/j.jaci.2012.02.023) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Comparison of allergen-specific IgG4 levels in CB and MB analyzed by the DLC chip. Allergen-specific IgG4 levels in paired samples of CB and MB (n = 92) depicted in BUg4. The cutoff value was 0.53 BUg4. A, α-Casein. B, Milk. C, Ovomucoid. Spearman's rank correlation test was used to assess the relation between CB and MB. Journal of Allergy and Clinical Immunology 2012 130, 113-121.e2DOI: (10.1016/j.jaci.2012.02.023) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions