Allergy Asthma Immunol Res.  2013 Nov;5(6):397-401. 10.4168/aair.2013.5.6.397.

The Effects of Storage Conditions on the Stability of House Dust Mite Extracts

• Jeong KY
• Choi SY
• Han IS
• Lee JH
• Lee JS
• Hong CS
• Park JW

• KMID: 2260284
• DOI: http://doi.org/10.4168/aair.2013.5.6.397

Abstract

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 -20degrees C, 4degrees C, 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, 4degrees C, and -20degrees C, 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 4degrees C regardless of the buffer conditions.
CONCLUSIONS
Storage temperature is the most important factor in preserving allergenicity of HDM extracts, which is ideal at 4degrees C. 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.

Keyword

Allergen; house dust mite; stability

MeSH Terms

Enzyme-Linked Immunosorbent Assay
Glycerol
Humans
Pyroglyphidae
Serum Albumin
Water
Glycerol
Serum Albumin
Water

Figure

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  Fig. 1 Changes in protein concentration in D. pteronyssinus extracts stored at room temperature (A), 4℃ (B), and -20℃ (C).

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  Fig. 2 Changes in Der p 1 concentration in D. pteronyssinus extracts stored at room temperature (A), 4℃ (B), and -20℃ (C).

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  Fig. 3 Changes in allergenicity of D. pteronyssinus extracts stored at room temperature (A) and 4℃ (B).

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  Fig. 4 SDS-PAGE profile (A) and IgE reactive bands (B) of D. pteronyssinus extracts reconstituted in distilled water and stored at 4℃. The numbers (0 to 52) indicate the duration (week) of storage after reconstitution. Arrow indicates the location of Der p 2.

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