Production and characterization of monoclonal antibodies to the steroid receptor coactivator AIB1
Production and characterization of monoclonal antibodies to the steroid receptor coactivator AIB1
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DOI：
10.1089/027245799315943
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发表时间：
1999-06-01
期刊：
影响因子：
--
通讯作者：
Meltzer, PS
中科院分区：
文献类型：
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作者：
Azorsa, DO;Meltzer, PS
类固醇辅活化子AIB1(在乳腺癌中扩增)是SRC-1反式激活辅活化子家族的成员，在大约7%的乳腺肿瘤样本中扩增。建立了分泌抗重组谷胱甘肽-S转移酶融合蛋白(GST-AIB.T1：A.A.)的杂交瘤细胞。605-1294)。从这些杂交瘤中分离出4株单抗，分别命名为AX15.1、AX15.2、AX15.3和AX15.4。Western印迹分析和酶联免疫吸附试验表明，4株单抗与AIB1特异性结合，不与两个类似的辅活化子SRC-1和TIF2发生交叉反应。4株单抗的Western印迹分析表明，这4株单抗在含有AIB1基因扩增的细胞裂解产物中特异性识别AIB1蛋白为160 kDa条带。这些单抗在体外翻译AIB1和代谢标记细胞中的AIB1。结果表明，这些新发现的单抗可用于研究AIB1的功能和表达。
The steroid coactivator AIB1 (Ampified In Breast cancer 1) is a member of the SRC-1 family of transactivation coactivators that is amplified in about 7% of breast tumor samples. Hybridomas were established that secreted monoclonal antibodies (MAbs) to a recombinant glutathione-S-transferase (GST) fusion protein expressing the steroid receptor interacting domain of AIB1 (GST-AIB.T1: a.a. 605-1294). Four MAbs from these hybridomas were characterized and designated AX15.1, AX15.2, AX15.3, and AX15.4. The MAbs were shown to be specific to AIB1 and did not cross-react with two similar coactivators SRC-1 and TIF2 as shown in Western blot analysis and enzyme-linked immunoadsorbent assay (ELISA), Western blot analysis using the four MAbs showed specific recognition of AIB1 protein as a 160 kDa band in lysates from cell lines containing AIB1 gene-amplification. The MAbs immunoprecipitated in vitro-translated AIB1 and cellular AIB1 from metabolically labeled cells. The results show that these newly described MAbs are useful in studies of AIB1 function and expression.