The stability of house dust mite allergens in glycerinated extracts - 04/09/11
Abstract |
Background: Mite allergen vaccines are important diagnostic and immunotherapeutic reagents. Previous studies on mite allergen stability under different storage conditions have yielded contradictory results. Objective: We sought to compare, over a 12-month period, the stability of mite allergens reconstituted in 50% glycerol and stored at different temperatures and to examine the role of protease inhibitors in enhancing allergen stability. Methods: Lyophilized allergen extracts were reconstituted in 50% glycerol, with and without protease inhibitors, and stored at –70°C, –20°C, 4°C, or 37°C for 12 months. At 6 and 12 months, the extracts were compared with freshly dissolved extracts by competition ELISA with pooled allergic sera, 2-site ELISA with mite-specific mAbs, and immunoblot analyses. Results: The overall potencies of the stored extracts measured by competition ELISA were stable at –20°C and 4°C. As determined by means of the immunoblot and 2-site ELISA, Der f 1 levels decreased at 4°C. Levels of Der f 2, Der p 1, and Der p 2 decreased in at least one of the allergen-specific assays. Storage at 37°C led to overall loss of potency and allergen content, whereas storage at –70°C was associated with a moderate loss of potency that increased with multiple freeze-thaw cycles. Protease inhibitors had no effect on allergen stability. Conclusion: Although overall potency of the extracts, as measured by competition ELISA, was preserved at –20°C and 4°C, allergen-specific assays indicated loss of allergens. These findings suggest that the competition ELISA is insensitive to decreases in the concentrations of individual allergens. (J Allergy Clin Immunol 2000;105:482-8.)
Il testo completo di questo articolo è disponibile in PDF.Keywords : Dust mite, monoclonal antibody, allergen, ELISA, immunoblot, stability, allergy, allergen extract
Abbreviations : CBER:, CI:, RP:
Mappa
Supported in part by an appointment (L.N.S.) to the Postgraduate Research Participation Program at the Center for Biologics Evaluation and Research administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the US Department of Energy and the US Food and Drug Administration. |
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The views expressed in this article are the personal opinions of the authors and are not the official opinion of the US Food and Drug Administration or the Department of Health and Human Services. |
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Reprint requests: Lyudmila N. Soldatova, PhD, Laboratory of Immunobiochemistry, FDA/CBER/DBPAP, 1401 Rockville Pike (HFM-422), Rockville, MD 20852. |
Vol 105 - N° 3
P. 482-488 - Marzo 2000 Ritorno al numeroBenvenuto su EM|consulte, il riferimento dei professionisti della salute.
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