Clarke J T, Embil J A
Biochim Biophys Acta. 1979 Jan 18;582(2):283-94. doi: 10.1016/0304-4165(79)90391-x.
Antiserum against galactosyl(alpha 1 leads to 4)galactosyl(beta 1 leads to 4)glucosylceramide (globotriaosylceramide, Gb3) was raised in rabbits by the administration of four weekly intramuscular injections of 1.5 mg of the purified glycoplipid along with bovine serum albumin and Freund's complete adjuvant. AntiGb3 activity was quantitated initially by immunoprescipitation employing Gb3 mixed with 100-fold excess of lecithin and cholesterol (1 : 1 or 1 : 2, by wt.) as antigen. Subsequently, complement fixation tests done with antigen preparations containing Gb3/lecithin/cholesterol (1 :6 :20, by wt.) showed antiGb3 titres of up to 1 : 8192. Fractionation of the antiserum by BioGel A5m chromatography indicated the antibody was an IgM immunoglobulin. The partially purified antibody stimulated complement-dependent release of glucose from glucose-containing liposomes prepared with shingomyelin/cholesterol/dicetylphosphate/Gb3 (molar ratio, 100 : 75 :11 :1). The antibody crossreacted with the trisaccharide, Gal(alpha1 leads to 4)Gal(beta1 leads to 4)Glc, but not with galactosylerceramide, lactosylceramide, GM1 ganglioside, globotetraosylceramide, digalactosyldiglyceride or a number of low molecular weight saccharides.
通过每周一次给兔子肌肉注射1.5毫克纯化糖脂，同时加入牛血清白蛋白和弗氏完全佐剂，连续注射四周，制备了抗半乳糖基（α1→4）半乳糖基（β1→4）葡萄糖神经酰胺（球三糖神经酰胺，Gb3）的抗血清。最初通过免疫沉淀法对抗Gb3活性进行定量，使用Gb3与100倍过量的卵磷脂和胆固醇（重量比为1:1或1:2）混合作为抗原。随后，用含有Gb3/卵磷脂/胆固醇（重量比为1:6:20）的抗原制剂进行补体固定试验，结果显示抗Gb3效价高达1:8192。通过BioGel A5m色谱法对抗血清进行分级分离表明，该抗体是一种IgM免疫球蛋白。部分纯化的抗体刺激了用鞘磷脂/胆固醇/二鲸蜡基磷酸酯/Gb3（摩尔比为100:75:11:1）制备的含葡萄糖脂质体中葡萄糖的补体依赖性释放。该抗体与三糖Gal（α1→4）Gal（β1→4）Glc发生交叉反应，但与半乳糖神经酰胺、乳糖神经酰胺、GM1神经节苷脂、球四糖神经酰胺、二半乳糖二甘油酯或多种低分子量糖类不发生交叉反应。
