APPLICATION OF LIPOSOMES TO GENERATION OF MONOCLONAL-ANTIBODY TO GLYCOSPHINGOLIPID - PRODUCTION OF MONOCLONAL-ANTIBODY TO GGOSE4CER
APPLICATION OF LIPOSOMES TO GENERATION OF MONOCLONAL-ANTIBODY TO GLYCOSPHINGOLIPID - PRODUCTION OF MONOCLONAL-ANTIBODY TO GGOSE4CER
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DOI：
10.1093/oxfordjournals.jbchem.a122041
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发表时间：
1987-07-01
影响因子：
2.7
通讯作者：
YASUDA, T
中科院分区：
文献类型：
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作者：
WATARAI, S;HANDA, S;YASUDA, T
将脂质体应用于GgOse4Cer的免疫和筛选GgOse4Cer的单克隆抗体的产生。用GgOse4Cer和明尼苏达沙门氏菌R595脂多糖掺入由二棕榈酰磷脂酰胆碱和胆固醇组成的脂质体对4周龄和22周龄的Balb/c小鼠进行免疫。由于免疫后 22 周龄 Balb/c 小鼠对 GgOse4Cer 的抗体反应高于 4 周龄 Balb/c 小鼠，因此在生成 GgOse4Cer 单克隆抗体之前使用 22 周龄 Balb/c 小鼠进行免疫。使用含有 GgOse4Cer 的脂质体，通过补体依赖性脂质体免疫裂解测定进行单克隆抗体的筛选。建立了IgM类的六种单克隆抗体：AG-1、-2、-3、-4、-5和-6。所获得的单克隆抗体的特异性通过使用掺入脂质体中的各种鞘糖脂的补体依赖性脂质体免疫裂解测定以及通过免疫染色的薄层色谱（TLC）来确定。在脂质体免疫裂解测定中，所有单克隆抗体仅与 GgOse4Cer 发生反应。此外，在TLC免疫染色中，单克隆抗体仅与GgOse4Cer发生反应。然而，所获得的单克隆抗体均不能在体外消除 C3H/He 小鼠脾细胞悬浮液的自然杀伤活性。脂质体可用于免疫和筛选过程中，以产生针对 GSL 的抗血清和单克隆抗体。
Liposomes were applied to the immunization with GgOse4Cer and screening for production of monoclonal antibody to GgOse4Cer. Four-week-old and 22-week-old Balb/c mice were immunized with GgOse4Cer and Salmonella minnesota R595 lipopolysaccharides incorporated liposomes which were composed of dipalmitoyl-phosphatidylcholine and cholesteroal. Since antibody response to GgOse4Cer was higher in 22-week-old than 4-week-old Balb/c mice after immunization, 22-week-old Balb/c mice were used for the immunization prior to generation of the monoclonal antibodies to GgOse4Cer. The screening of monoclonal antibodies were performed by complement-dependent liposome immune lysis assay using GgOse4Cer-containing liposomes. Six kinds of monoclonal antibodies, AG-1, -2, -3, -4, -5, and -6, of the IgM class were established. The specificities of the monoclonal antibodies obtained were defined by complement-dependent liposome immune lysis assay using various glycosphingolipids incorporated in liposomes and by thin-layer chromatography (TLC) with immunostaining. All of the monoclonal antibodies reacted only with GgOse4Cer in the liposome immune lysis assay. In addition, the monoclonal antibodies reacted only with GgOse4Cer in the TLC immunostaining. However, none of the monoclonal antibodies obtained was capable of removing natural killer activity from C3H/He mice spleen cell suspensions in vitro. Liposomes may be useful in the procedures of immunization and screening for generation of antiserum and monoclonal antibody to GSLs.