Stability of extracts from pollens of allergenic importance in Korea

BACKGROUND/AIMS: Accurate diagnosis and the effects of allergen-specific immunotherapy for pollinosis are greatly dependent on the potency and stability of the extract. This study aimed to examine factors, such as temperature and storage buffer composition, that affect the stability of allergen extracts from pollens of allergenic importance in Korea. METHODS: We prepared four pollen allergen extracts from ragweed, mugwort, Japanese hop, and sawtooth oak, which are the most important causes of seasonal rhinitis in Korea. Changes of protein and major allergen concentration were measured over 1 year by Bradford assay, two-site enzyme-linked immunosorbent assay, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis after reconstitution of the lyophilized allergen extract in various buffers and stored at room temperature (RT, 18°C to 26°C) or refrigerated (4°C). RESULTS: More than 90% of the original protein concentration in all four extracts examined was detected over 1 year when 50% glycerol was added and refrigerated, whereas 57.9% to 94.5% remained in the extracts at RT. The addition of 50% glycerol to the storage buffer was found to prevent protein degradation at RT. Amb a 1, a major allergen of ragweed, was almost completely degraded in 9 weeks at RT when reconstituted in a buffer without 50% glycerol. However, 55.6% to 92.8% of Amb a 1 content was detected after 1 year of incubation at 4°C in all buffer conditions except 0.3% phenol. CONCLUSION: Addition of 50% glycerol as well as refrigeration was found to be important in increasing the shelf-life of allergen extracts from pollens of allergenic importance.ope

Optimal conditions for the storage of German cockroach extract

Allergen extracts are commonly utilized for diagnosis and immunotherapy; however, the stability of protease‑rich extracts is important for a precise diagnosis and treatment efficacy. The present study determines the optimal conditions for the storage of German cockroach allergen extract. Cockroach extracts were reconstituted in four buffers: normal saline (NS), 50% glycerol in NS, 0.3% phenol in NS, or 0.3% phenol and 50% glycerol in NS. The extracts in different buffers were stored either at room temperature (18‑26˚C, RT) or refrigerated (2‑8˚C). Subsequently, the protein concentration and allergen content (Bla g 1 and Bla g 2) in the extracts were examined for the course of one year. Extract potency was estimated by inhibition ELISA. At least 90.5% protein, 94.4% Bla g 1, 65.2% Bla g 2, and 91.4% potency remained after one year when 50% glycerol NS was added to the extract with refrigeration. However, less than 13.7% protein, 17.1% Bla g 1, 0% Bla g 2 and 32.5% potency were maintained after one year when 50% glycerol NS was not added to the extract and was maintained at RT. The addition of 0.3% phenol NS did not show significant effects on extract stability. The addition of 50% glycerol NS and refrigerated storage temperature were found to be important factors for increasing the shelf life of protease‑rich cockroach extract.restrictio

Method for diagnosing and screening asthma using Neuropilin １

본 발명은 뉴로필린(Neuropilin, NRP) 유전자를 포함하는 천식 진단 및 스크리닝용 키트 및 이를 이용한 천식 치료제 스크리닝 방법에 관한 것으로서, 보다 구체적으로 천식 환자 및 집먼지진드기 추출물에 의해서 NRP1의 발현이 증가되며, NRP1의 발현 억제에 의해서 천식과 관련된 MMP9의 mRNA, 단백질 발현, 효소 활성 및 프로모터의 활성이 감소하는 결과를 통해서 NRP1을 천식 치료제 개발을 위한 표적으로서 유용하게 사용될 수 있음을 확인하였다.ope