Klysner S, Welinder K G, Løwenstein H, Matthiesen F
ALK Research, Hørsholm, Denmark.
Clin Exp Allergy. 1992 Apr;22(4):491-7. doi: 10.1111/j.1365-2222.1992.tb00152.x.
Monoclonal antibodies (PpV4) raised against Phleum pratense group V allergen were used for immuno-affinity chromatography of cross-reacting group V allergens from related grass species. Fractions enriched in group V allergen were obtained from Lolium perenne, Poa pratense and Dactylis glomerata extracts. The major components in these fractions were found in the Mwr range 25-28 kD. IgE binding to these components was shown using a pool of grass allergic sera, by SDS-PAGE immunoblotting. These fractions were electroblotted from tricine SDS-PAGE gels onto a polyvinylidene-difluoride membrane and selected group V bands were directly cut out and used for amino acid analysis and NH2-terminal sequencing. Both the amino acid compositions and the NH2-terminal sequences obtained for each group V allergen were almost similar to each other and to the sequence and composition of the previously described allergen Phl p V from Phleum pratense. A common trait of the investigated allergens, is the very high contents of alanine (25-32%) and the presence of the modified amino acid, hydroxyproline.
针对梯牧草V组变应原产生的单克隆抗体（PpV4）用于对相关禾本科物种中交叉反应性V组变应原进行免疫亲和层析。从多年生黑麦草、草地早熟禾和鸭茅提取物中获得了富含V组变应原的级分。这些级分中的主要成分分子量范围在25 - 28 kD。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳免疫印迹法，使用一组对草过敏的血清证明了免疫球蛋白E与这些成分的结合。将这些级分从三羟甲基氨基甲烷 - 十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳凝胶电印迹到聚偏二氟乙烯膜上，直接切下选定的V组条带用于氨基酸分析和氨基末端测序。每个V组变应原获得的氨基酸组成和氨基末端序列彼此几乎相似，并且与先前描述的梯牧草Phl p V变应原的序列和组成相似。所研究变应原的一个共同特征是丙氨酸含量非常高（25 - 32%）以及存在修饰氨基酸羟脯氨酸。
