Bouhours J F, Richard C, Ruvoen N, Barreau N, Naulet J, Bouhours D
Institut de Transplantation et de Recherche en Transplantation, INSERM Unité 437, Centre Hospitalier Universitaire, Nantes, France.
Glycoconj J. 1998 Jan;15(1):93-9. doi: 10.1023/a:1006903919461.
A polyclonal antibody was raised against the Galalpha1-3Gal carbohydrate epitope, which is expressed by all mammals (except man and the closest primate species) by immunizing hens with rabbit erythrocyte membranes. IgY was isolated from egg yolks, and affinity-purified on a Galalpha1-3Gal-Synsorb column. Two percent of the initial IgY fraction was recovered. The specificity of the affinity-purified antibody was characterized by: absorption with human, rabbit and pig erythrocytes; by using Synsorb columns; by inhibition with different saccharides; and by immunostaining of glycolipids separated on thin layer chromatograms. A weak reactivity was found toward blood group B or blood group Pk determinant, depending on the assay system used. Such reactivities were abolished after absorption by the appropriate sorbents, yielding a polyclonal anti-Galalpha1-3Gal antibody with narrow specificity.
通过用兔红细胞膜免疫母鸡,制备了一种针对α-半乳糖基-1,3-半乳糖碳水化合物表位的多克隆抗体,该表位由所有哺乳动物(除人类和与其亲缘关系最近的灵长类物种外)表达。从蛋黄中分离出IgY,并在α-半乳糖基-1,3-半乳糖-Synsorb柱上进行亲和纯化。回收了初始IgY组分的2%。亲和纯化抗体的特异性通过以下方法进行表征:用人、兔和猪红细胞进行吸收;使用Synsorb柱;用不同糖类进行抑制;以及对在薄层色谱上分离的糖脂进行免疫染色。根据所使用的检测系统,发现对B血型或Pk血型决定簇有弱反应性。在用适当的吸附剂吸收后,这些反应性消失,从而产生了具有狭窄特异性的多克隆抗α-半乳糖基-1,3-半乳糖抗体。