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Allergy Asthma Immunol Res. 2013 Nov;5(6):397-401. English.
Published online 2013 July 19. http://dx.doi.org/10.4168/aair.2013.5.6.397 |
| Copyright © 2013 The Korean Academy of Asthma, Allergy and Clinical Immunology • The Korean Academy of Pediatric Allergy and Respiratory Disease |
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Kyoung Yong Jeong,1 Soo-Young Choi,1 In-Soo Han,1 Jae-Hyun Lee,1 Joo-Shil Lee,2 Chein-Soo Hong,1 andJung-Won Park 1 |
| 1Departmet of Internal Medicine and Institute of Allergy, Yonsei University College of Medicine, Seoul, Korea. |
| 2Center for Immunology and Pathology, Korea National Institute of Health, Seoul, Korea. |
Correspondence to: Jung-Won Park, MD, PhD, Department of Internal Medicine, Institute of Allergy, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Korea. Tel: +82-2-2228-1961; Fax: +82-2-362-8647; Email: parkjw@yuhs.ac
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Received December 04, 2012; Revised January 25, 2013; Accepted January 31, 2013.
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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Abstract
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Purpose
Allergen extracts from the house dust mite (HDM, Dermatophagoides pteronyssinus) are widely utilized for diagnosis and treatment of allergic diseases. It is known that allergen extracts degrade and lose potency when stored over time.
Methods
This study aimed to determine the optimal conditions for stability of allergen extracts. This study was undertaken to investigate the optimal storage conditions for HDM extracts, the effects of adding 0.03% human serum albumin (HSA) and 50% glycerol were evaluated at -20℃, 4℃, and room temperature (RT). Changes in protein and group 1 major allergen (Der p 1) concentration, as well as allergenicity were measured over a 1 year period using the Bradford assay, two-site ELISA, and ELISA inhibition.
Results
Protein concentrations decreased by 86%, 51%, and 6% at RT, 4℃, and -20℃, respectively, when stored in distilled water. Overall allergenicity remained high (89.9%) when the extracts was reconstituted in 50% glycerol solution, and was 93.1% when reconstituted in 50% glycerol and 0.03% HSA at RT. Allergenicity was decreased to 36.6% and 33.3%, however, reconstitution in DW or 0.03% HSA solution at RT, respectively. Allergenicity was remained high as 92.0%-97.0% when stored at 4℃ regardless of the buffer conditions.
Conclusions
Storage temperature is the most important factor in preserving allergenicity of HDM extracts, which is ideal at 4℃. The addition of 50% glycerol to the storage buffer was also found to play an important role in increasing the shelf-life of HDM extracts at RT.
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Keywords: Allergen, house dust mite, stability.
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Open Access
Article
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