Ventas P, Carreira J, Polo F
Research Department, Alergia e Inmunología Abelló S.A., Madrid, Spain.
Immunol Lett. 1991 Aug;29(3):229-34. doi: 10.1016/0165-2478(91)90175-a.
The allergen composition of one of the most important storage mites, Lepidoglyphus destructor, has been studied by immunodetection after SDS-PAGE with individual patient sera. An allergenic polypeptide of 14 kDa was identified with 95% of the sera. This major allergen was isolated in the supernatant of 60% ammonium sulfate salt precipitation of the whole extract, which was subsequently used to immunize BALB/c mice so as to produce monoclonal antibodies. Four mAbs recognizing molecules with IgE-binding ability were obtained. The specificity of the mAbs was assayed against different allergenic extracts, and the molecules recognized by them were characterized by immunoblotting. Two mAbs (Le5B5 and Le9E4) were directed to the 14-kDa allergen; the other two to several proteins of lesser allergenic significance.
利用个体患者血清,通过SDS - PAGE后的免疫检测法,对最重要的仓储螨之一——毁灭嗜鳞螨的变应原成分进行了研究。95%的血清可识别出一条14 kDa的变应原多肽。该主要变应原在全提取物60%硫酸铵盐沉淀的上清液中分离得到,随后用于免疫BALB/c小鼠以产生单克隆抗体。获得了4种识别具有IgE结合能力分子的单克隆抗体。针对不同的变应原提取物检测了这些单克隆抗体的特异性,并通过免疫印迹法对它们识别的分子进行了表征。两种单克隆抗体(Le5B5和Le9E4)针对14 kDa的变应原;另外两种针对变应原性较低的几种蛋白质。