Boluda L, Casanovas M, Prieto J L, Fernández-Caldas E
C.B.F. LETI, S.A. Research & Development Laboratories, Tres Cantos, Madrid, Spain.
J Investig Allergol Clin Immunol. 1998 Jul-Aug;8(4):207-13.
Par j 1 is the major allergen of Parietaria judaica. The objectives of this study were the following: 1) to purify Par j 1; 2) to develop an enzyme immunoassay based on the bivalent properties of specific IgE and IgG to determine the Par j 1 content in several batches of P. judaica extracts; and, 3) to study the contribution of Par j 1 to the total allergenicity and antigenicity of P. judaica extracts. P. judaica pollen was extracted and subjected to hydrophobic interaction and gel filtration chromatography for the purification of Par j 1. Inhibition enzyme immunoassays, SDS-PAGE and immunoblotting were used to characterize the allergen content. The in vivo biological potency of the extracts was estimated by skin prick testing 26 P. judaica clinically sensitive patients. The new enzyme immunoassay showed a high degree of specificity and sensitivity, detecting from 2 to 100 ng Par j 1/ml. The range of Par j 1 content in nine batches ranged from 23% to 78% of the total protein in the extracts. The Par j 1 content showed a significant correlation with the allergenic potency of these extracts evaluated by specific IgE inhibition and skin prick testing; the correlation with the specific IgG inhibition capacity was not significant. Purified Par j 1 shows great specific IgE and IgG binding capacity; its content can be determined using this newly developed enzyme immunoassay. Par j 1 levels exhibit a significant correlation with the biological potency of the extracts. This method allows the detection of Par j 1 isoforms.
Par j 1是墙草（Parietaria judaica）的主要变应原。本研究的目的如下：1）纯化Par j 1；2）基于特异性IgE和IgG的二价特性开发一种酶免疫测定法，以测定几批墙草提取物中的Par j 1含量；3）研究Par j 1对墙草提取物总变应原性和抗原性的贡献。提取墙草花粉，并进行疏水相互作用和凝胶过滤色谱以纯化Par j 1。采用抑制酶免疫测定法、SDS-PAGE和免疫印迹法对变应原含量进行表征。通过对26例对墙草临床敏感的患者进行皮肤点刺试验来评估提取物的体内生物学效价。新的酶免疫测定法显示出高度的特异性和敏感性，可检测出2至100 ng Par j 1/ml。九批提取物中Par j 1的含量范围为提取物总蛋白的23%至78%。Par j 1含量与通过特异性IgE抑制和皮肤点刺试验评估的这些提取物的变应原效价呈显著相关；与特异性IgG抑制能力的相关性不显著。纯化的Par j 1显示出很强的特异性IgE和IgG结合能力；其含量可以使用这种新开发的酶免疫测定法来测定。Par j 1水平与提取物的生物学效价呈显著相关。该方法能够检测Par j 1的异构体。
