Monoclonal antibodies to Escherichia coli 50S ribosomes.
Monoclonal antibodies to Escherichia coli 50S ribosomes.
复制标题
大肠杆菌 50S 核糖体单克隆抗体。
DOI：
10.1093/nar/8.20.4639
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发表时间：
1980
影响因子：
14.9
通讯作者：
Daugherty,B
中科院分区：
文献类型：
--
作者：
Shen,V;King,TC;Kumar,V;Daugherty,B
已分离出产生针对5os核糖体蛋白的单抗的杂交瘤细胞系。将从Escherichiacoli中提取的50S核糖体亚基免疫的BALB/c小鼠的脾细胞与小鼠骨髓瘤细胞系SP2/0-Ag14融合，用双抗体夹心法对产生抗体的杂交瘤细胞进行初步筛选，然后在软琼脂中克隆杂交瘤细胞。抗体的特征是它们与分离在磷酸纤维素胶和二维聚丙烯酰胺凝胶上的单个50S核糖体蛋白具有特异性结合。用纯化的单个核糖体蛋白证实了这些指认。到目前为止分析的四个克隆中，有两个与r-蛋白L5的特异性相同。其他克隆产生两种针对r-蛋白L20的不同抗体。每个单抗形成核糖体二聚体，在电子显微镜下可见。二聚体可以与不同的第二抗体反应，形成包含8个或更多核糖体的链，这可能对结构研究有用。
Hybridoma cell lines that produce monoclonal antibodies directed against 5OS ribosomal proteins have been isolated. Spleen cells (from BALB/c mice immunized with 50S ribosomal subunits extracted fromEscherichiacoli) were fused to mouse myeloma cell line SP2/0-Ag 14. The initial screening for antibody producing hybridomas was carried out by a double antibody sandwich method; hybridomas were subsequently cloned in soft agar. Antibodies were characterized by their specific binding to individual 50S ribosomal proteins separated on phosphocellulose cohmins and in two-dimensional polyacrylamide gels. The assignments were confirmed with purified single ribosomal proteins. Of four clones analyzed thus far, two are identical with specificity for r-protein L5. The other clones produce two different antibodies directed against r-protein L20. Each monoclonal antibody formed ribosome dimers visualizable in the electron microscope. Dimers could be reacted with a different second antibody to form chains containing 8 or more ribosomes, which may be useful for structural studies.