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Characterization of immunoglobulin E-binding proteins (IgE) of scomberomorus commerson lacepede

机译:鼠免疫球蛋白E结合蛋白(IgE)的表征

摘要

The objective of this study was to determine the Immunoglobulin E-binding proteins (IgE) and major allergens of Scomberomorus commerson Lacepede(Narrow-barred Spanish mackerel). Allergen extracts were obtained from uncooked and cooked fish by homogenization in phosphate-buffered saline followed by continuous extraction at 4oC or on ice. Protein profiles and IgEbinding patterns were then detected by means of sodium dodecyl polyacrylamidegel electrophoresis (SDS-PAGE) and immunoblotting using sera from patients sensitized to the fish. SDS-PAGE of the uncooked fish extracts revealed 26 protein bands in the range of about 11 to >175 kD, while the cooked extracts produced fewer protein bands. Immunoblotting demonstrated 17 IgE-binding bands, ranging in molecular weight from 11 to 151 kD. Two components with molecular weight of about ~50 and 42 kD showed the highest frequency of IgE-bindingud(62.2 and 51.4% respectively) and were identified as the major allergens of this fish allergy. Other IgE-binding proteins including a protein at ~12 kD which was equivalent in size to parvalbumin were identified as the minorudallergens.
机译:这项研究的目的是确定Scomberomorus commerson Lacepede(窄条鲭鱼)的免疫球蛋白E结合蛋白(IgE)和主要过敏原。从未煮熟和煮熟的鱼中提取过敏原,方法是在磷酸盐缓冲液中匀浆,然后在4℃或冰上连续提取。然后通过十二烷基聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹法(使用对鱼敏感的患者的血清)检测蛋白谱和IgE结合模式。未煮过的鱼提取物的SDS-PAGE显示26条蛋白带,范围在11至> 175 kD之间,而煮熟的提取物产生的蛋白带较少。免疫印迹显示17条IgE结合带,分子量在11至151 kD之间。分子量约为50kD和42kD的两个成分显示出最高的IgE结合频率(分别为62.2%和51.4%),并被确定为该鱼过敏的主要过敏原。其他IgE结合蛋白,包括大小约等于小白蛋白的约12 kD蛋白,被鉴定为次要变应原。

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