The stability of house dust mite allergens in glycerinated extracts

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The stability of house dust mite allergens in glycerinated extracts Lyudmila N. Soldatova, PhD, Elizabeth J. Paupore, BS, Suzann H. Burk, BA, Richard W. Pastor, PhD, Jay E. Slater, MD Journal of Allergy and Clinical Immunology Volume 105, Issue 3, Pages 482-488 (March 2000) DOI: 10.1067/mai.2000.104549 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 The RPs of stored mite extracts were determined by means of competition ELISA with pooled allergic sera, comparing the stored extracts to freshly reconstituted reference material (RP, 1.0). The storage temperatures were –70°C, –20°C, 4°C, and 37°C. Bars represent the average values of RP for 3 parallel ELISA plates. Error bars indicate 95% CIs of each set of data. The 95% CI of reference extract ranges from 0.699 to 1.43 when N = 3. Journal of Allergy and Clinical Immunology 2000 105, 482-488DOI: (10.1067/mai.2000.104549) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Immunoblots of mite extracts separated by SDS-PAGE on a 16% polyacrylamide gel, transferred to a nitrocellulose membrane, and developed with pooled allergic sera (6-month study). Storage temperatures were –20°C, 4°C, and 37°C. A shows extract JE 318, and B shows extract JE 312. L, Lyophilized extract; +, extracts with added protease inhibitors; –, extracts without added protease inhibitors. Journal of Allergy and Clinical Immunology 2000 105, 482-488DOI: (10.1067/mai.2000.104549) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 Group 1 and group 2 allergen content of mite extracts was determined by using 2-site ELISA with mono-clonal capturing antibodies and biotinylated monoclonal detection antibodies (6-month study). Storage temperatures were –20°C, 4°C, and 37°C. Bars represent the average values of allergen content of extracts incubated at different temperatures in multiple parallel microtiter plates; error bars indicate 95% CIs based on a pooled SD of 17 μg/mL. Journal of Allergy and Clinical Immunology 2000 105, 482-488DOI: (10.1067/mai.2000.104549) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 RPs of stored mite extracts were compared with allergenspecific RPs, which were calculated by dividing the allergen content of the stored extract by the allergen content of the freshly reconstituted control. Six-month and 12-month data are shown. Journal of Allergy and Clinical Immunology 2000 105, 482-488DOI: (10.1067/mai.2000.104549) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 Immunoblots of D farinae mite extracts separated by SDS-PAGE on 16% polyacrylamide gel, transferred to a polyvinylene difluoride membrane, and developed with anti-group 1 (A) and anti-group 2 (B) mAbs (12-month study). Storage temperatures were –70°C, –20°C, 4°C, and 37°C. L, Lyophilized extract. Journal of Allergy and Clinical Immunology 2000 105, 482-488DOI: (10.1067/mai.2000.104549) Copyright © 2000 Mosby, Inc. Terms and Conditions