Leduc-Brodard V, Inacio F, Jaquinod M, Forest E, David B, Peltre G
Unité d'Immuno-Allergie, Institut Pasteur, Paris, France.
J Allergy Clin Immunol. 1996 Dec;98(6 Pt 1):1065-72. doi: 10.1016/s0091-6749(96)80193-x.
Monoclonal antibodies were produced against Dac g 4, a purified major basic allergen from Dactylis glomerata pollen. Their ability to be used for immunopurification of Dac g 4 was studied on a BIAcore apparatus (Pharmacia). The allergen was purified by affinity chromatography with one monoclonal antibody. Its precise molecular mass, 59,185 +/- 30 d, was determined by mass spectrometry. Its isoelectric point is 10.4. Sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting showed that Dac g 4-related proteins of similar molecular mass were detected in the majority of allergenic grass pollen species. By double-site ELISAs, we have estimated that Dac g 4 represents about 6% of the total proteins from a water-soluble extract. One monoclonal antibody (mAb H) recognized a 60 kd cross-reactive protein in other grass pollens, though none in any of the tree or weed pollens tested. Inhibition studies of IgE antibody binding to Dac g 4 with pollen extracts confirmed the presence of cross-reactive allergens in Secale cereale, Lolium perenne, Festuca elatior, Holcus lanatus, Bromus arvensis, Poa pratense, Hordeum sativum, and Phleum pratense.
制备了针对Dac g 4的单克隆抗体，Dac g 4是从鸭茅花粉中纯化得到的一种主要碱性变应原。在BIAcore仪器（Pharmacia公司）上研究了其用于免疫纯化Dac g 4的能力。用一种单克隆抗体通过亲和层析法纯化变应原。通过质谱法测定其精确分子量为59,185±30 d。其等电点为10.4。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹显示，在大多数变应性禾本科花粉物种中检测到了分子量相似的Dac g 4相关蛋白。通过双位点酶联免疫吸附测定法，我们估计Dac g 4占水溶性提取物总蛋白的约6%。一种单克隆抗体（单克隆抗体H）在其他禾本科花粉中识别出一种60 kd的交叉反应蛋白，但在所测试的任何树木或杂草花粉中均未识别出。用花粉提取物对IgE抗体与Dac g 4结合的抑制研究证实，在黑麦、多年生黑麦草、高羊茅、绒毛草、野燕麦、草地早熟禾、栽培大麦和梯牧草中存在交叉反应性变应原。
