Suzuki Y, Nishi H, Hidari K, Hirabayashi Y, Matsumoto M, Kobayashi T, Watarai S, Yasuda T, Nakayama J, Maeda H
Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Science.
J Biochem. 1991 Feb;109(2):354-60.
A new monoclonal antibody (NS24) directed to the N-acetylneuraminyl alpha 2-3Gal beta 1-4GlcNAc residue in type II sugar chain of N-acetylneuraminyllactoneotetraosylceramide [sialylparagloboside, IV3(NeuAc)nLc4Cer] was prepared by hybridoma technique. Liposomes composed of dipalmitoylphosphatidylcholine, cholesterol, IV3(NeuAc)nLc4Cer, and lipopolysaccharides from Salmonella minnesota R595 were used for immunization with IV3(NeuAc)nLc4Cer isolated from human erythrocytes. This method allowed the fusion of spleen cells of immunized mouse with myeloma cells only three days after immunization. NS24 reacted specifically to both naturally occurring and chemically synthesized IV3-(NeuAc)nLc4Cer, whereas it has no reactivity to structurally related gangliosides, such as IV6(NeuAc)nLc4Cer, N-glycolylneuraminyl alpha 2-3lactoneotetraosylceramide [IV3(NeuGc)-nLc4Cer], i-active ganglioside [VI3(NeuAc)nLc6Cer], I-active ganglioside [VIII3(NeuAc)-VI3(NeuAc)IV6kladoLc8Cer], GM4(NeuAc), GM3(NeuAc), GM3(NeuGc), GM1b(NeuAc), GD3-(NeuAc), other ganglio-series gangliosides, sulfatide, and paragloboside (nLc4Cer). Synthetic N-acetylneuraminyl alpha 2-3lactotetraosylceramide [IV3(NeuAc)Lc4Cer] and its asialo-derivative (Lc4Cer) carrying type I sugar chain also showed no reaction with NS24. One to 100 pmol of IV3(NeuAc)nLc4Cer was detected dose-dependently by a thin-layer chromatography/enzyme immunostaining procedure. Human gastric carcinomas showed positive reactions with NS24 immunochemically and histochemically. NS24 reacted preferentially with poorly differentiated adenocarcinomas rather than well differentiated ones.
通过杂交瘤技术制备了一种新的单克隆抗体（NS24），该抗体针对N-乙酰神经氨酰乳糖新四糖神经酰胺[唾液酸副球蛋白苷，IV3(NeuAc)nLc4Cer] II型糖链中的N-乙酰神经氨酰α2-3Galβ1-4GlcNAc残基。由二棕榈酰磷脂酰胆碱、胆固醇、IV3(NeuAc)nLc4Cer和来自明尼苏达沙门氏菌R595的脂多糖组成的脂质体，用于用人红细胞中分离的IV3(NeuAc)nLc4Cer进行免疫。该方法使得免疫小鼠的脾细胞与骨髓瘤细胞仅在免疫三天后就能融合。NS24对天然存在的和化学合成的IV3-(NeuAc)nLc4Cer均有特异性反应，而对结构相关的神经节苷脂无反应，如IV6(NeuAc)nLc4Cer、N-羟乙酰神经氨酰α2-3乳糖新四糖神经酰胺[IV3(NeuGc)-nLc4Cer]、i-活性神经节苷脂[VI3(NeuAc)nLc6Cer]、I-活性神经节苷脂[VIII3(NeuAc)-VI3(NeuAc)IV6kladoLc8Cer] GM4(NeuAc)、GM3(NeuAc)、GM(NeuGc)、GM1b(NeuAc)、GD3-(NeuAc)、其他神经节系列神经节苷脂、硫脂和副球蛋白苷（nLc4Cer）。携带I型糖链的合成N-乙酰神经氨酰α2-3乳糖四糖神经酰胺[IV(NeuAc)Lc4Cer]及其去唾液酸衍生物（Lc4Cer）与NS24也无反应。通过薄层色谱/酶免疫染色法可剂量依赖性地检测到1至100 pmol的IV3(NeuAc)nLc4Cer。人胃癌在免疫化学和组织化学上与NS24呈阳性反应。NS24优先与低分化腺癌反应，而不是高分化腺癌。
