Varjonen E, Björkstén F, Savolainen J
Department of Dermatology, Helsinki University Central Hospital, Finland.
Clin Exp Allergy. 1996 Apr;26(4):436-43.
Ingested cereals and inhaled cereal dusts can cause IgE-mediated allergy. Testing for these allergies yields variable, often undependable results, which could be due to the use of unsuitable or degraded testing materials.
We studied the effect of storage media, time and temperature on the stability of protein allergens extracted from wheat, rye, barley and oats flour. We also examined the effect of heat processing similar to that used when preparing foods on the stability of allergenic proteins present in wheat flour.
After storage experiments proteins in allergen extracts were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then visualized using immunoblotting or staining with Coomassie brilliant blue. After heat processing wheat flour proteins were extracted and their IgE-binding capacity was estimated.
Extracted proteins from wheat, rye and barley flour appeared stable for at least 21 months when stored at 4 degrees C in an aqueous solution containing 50% glycerol and 0.45% sodium chloride. In a medium with 0.4% phenol and 0.9% sodium chloride degradation of many proteins became apparent already after 1 month of storage. Oats extracts lost their IgE-binding capacity more rapidly and the maximal storage time appeared to be 10 months at 4 degrees C in a solution containing 50% glycerol and 0.45% sodium chloride. The IgE-binding capacity of the allergenic proteins decreased as heat-processing temperature and heating time increased. No processing conditions entirely abolished IgE-binding.
Cereal allergen preparations can be stored either in lyophilized form or at 4 degrees C in 50% glycerol. Unheated materials should be used also when testing for food allergy, as heating reduces allergenicity and thus sensitivity at testing.
摄入谷物和吸入谷物粉尘可引发IgE介导的过敏反应。针对这些过敏的检测结果多变,往往不可靠,这可能是由于使用了不合适或已降解的检测材料。
我们研究了储存介质、时间和温度对从小麦、黑麦、大麦和燕麦粉中提取的蛋白质过敏原稳定性的影响。我们还研究了类似于食品制备过程中的热处理对小麦粉中存在的致敏蛋白稳定性的影响。
在储存实验后,使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离过敏原提取物中的蛋白质,然后通过免疫印迹或考马斯亮蓝染色进行可视化。热处理后,提取小麦粉蛋白并评估其IgE结合能力。
从小麦、黑麦和大麦粉中提取的蛋白质,当在含有50%甘油和0.45%氯化钠的水溶液中于4℃储存时,至少21个月内似乎是稳定的。在含有0.4%苯酚和0.9%氯化钠的介质中,许多蛋白质在储存1个月后就明显降解。燕麦提取物更快地失去其IgE结合能力,在含有50%甘油和0.45%氯化钠的溶液中于4℃下,最大储存时间似乎为10个月。致敏蛋白的IgE结合能力随着热处理温度和加热时间的增加而降低。没有任何加工条件能完全消除IgE结合。
谷物过敏原制剂可以冻干形式储存,或在50%甘油中于4℃储存。在进行食物过敏检测时也应使用未加热的材料,因为加热会降低致敏性,从而降低检测时的敏感性。