Gathuru J K, Higashi H, Kato S, Usuba O, Naiki M
Jpn J Vet Res. 1989 Apr;37(2):71-83.
An improved enzyme-linked immunosorbent assay (ELISA) for detection of heterophile Hanganutziu-Deicher (HD) antibodies and antigens, which are frequently detected in sera and/or cancerous tissues from patients with various cancers was developed using biotinylated chicken anti-GM3(NeuGc) antibody and avidin-horseradish peroxidase conjugate. The N-glycolylneuraminyllactosyl-ceramide, GM3(NeuGc) ganglioside was purified from horse erythrocyte membranes. The ELISA procedure required 300 ng GM3(NeuGc) antigen to coat plastic microtiter plates and 190 ng biotinylated antibody per well to give optimum product formation. The technique could detect 6 ng antigen in tissue homogenate as compared to 0.6 ng of the pure compound by inhibition. Chicken anti-GM3(NeuGc) antibody quantitatively inhibited the biotinylated antibody, however, this procedure was not suitable to quantify lower affinity HD antibody in patient sera. Immunostaining specific for HD antigen-positive cells, in tissue sections was by 4 micrograms/ml biotinylated antibody and 200 dilution of Avidin-biotinylated peroxidase complex reagent using pig intestine and lymph node as positive tissues and chicken intestine and lung as negative tissues.
利用生物素化鸡抗GM3（NeuGc）抗体和抗生物素蛋白-辣根过氧化物酶缀合物，开发了一种改进的酶联免疫吸附测定（ELISA），用于检测嗜异性汉加纽齐乌-戴歇尔（HD）抗体和抗原，这些抗体和抗原在各种癌症患者的血清和/或癌组织中经常被检测到。从马红细胞膜中纯化了N-糖基神经氨酰乳糖基神经酰胺，即GM3（NeuGc）神经节苷脂。ELISA程序需要300 ng GM3（NeuGc）抗原包被塑料微量滴定板，每孔需要190 ng生物素化抗体以实现最佳产物形成。与通过抑制法检测0.6 ng纯化合物相比，该技术能够检测组织匀浆中6 ng的抗原。鸡抗GM3（NeuGc）抗体可定量抑制生物素化抗体，然而，该程序不适用于定量患者血清中亲和力较低的HD抗体。在组织切片中，针对HD抗原阳性细胞的免疫染色采用4微克/毫升生物素化抗体和抗生物素蛋白-生物素化过氧化物酶复合物试剂200倍稀释，以猪肠和淋巴结作为阳性组织，鸡肠和肺作为阴性组织。
