Yadzir Zailatul Hani Mohamad, Misnan Rosmilah, Abdullah Noormalin, Bakhtiar Faizal, Arip Masita, Murad Shahnaz
Allergy and Immunology Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia.
Asian Pac J Trop Biomed. 2012 Jan;2(1):50-4. doi: 10.1016/S2221-1691(11)60189-5.
To characterize the major allergens of Macrobrachium rosenbergii (giant freshwater prawn).
Raw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools.
SDS-PAGE of the raw extract showed 23 protein bands (15-250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients' sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase.
It can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies.
鉴定罗氏沼虾（淡水大对虾）的主要过敏原。
制备淡水大对虾的生提取物和熟提取物。采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹技术，用20例皮肤点刺试验（SPT）阳性患者的血清鉴定IgE反应模式。然后，使用蛋白质组学方法，结合二维（2-DE）电泳、质谱和生物信息学工具，对鉴定出的主要过敏原进行表征。
生提取物的SDS-PAGE显示有23条蛋白带（15 - 250 kDa），但在熟提取物中未发现40至100 kDa范围内的蛋白带。免疫印迹实验表明，生提取物和熟提取物分别有11种和5种IgE结合蛋白，分子量范围为15至155 kDa。一种耐热的36 kDa蛋白被鉴定为两种提取物的主要过敏原。此外，一种42 kDa的热敏蛋白被证明是生提取物的主要过敏原。二维凝胶将对虾蛋白分离成50多个不同的蛋白点。其中，10个点与患者血清显示出特异性IgE反应。基质辅助激光解吸/电离飞行时间（MALDI-TOF）分析鉴定出2种重要过敏原，原肌球蛋白和精氨酸激酶。
可以得出结论，这些过敏原的可得性将有助于对虾过敏的基于成分的诊断和治疗。
