Masking of nontumorous antigens for development of human tumor nucleolar antibodies with improved specificity.
Masking of nontumorous antigens for development of human tumor nucleolar antibodies with improved specificity.
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发表时间：
1985-11
期刊：
影响因子：
11.2
通讯作者：
James W. Freeman;Rose K. Busch;B. E. Ross;Harris Busch
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文献类型：
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作者：
James W. Freeman;Rose K. Busch;B. E. Ross;Harris Busch
从HeLa细胞中纯化的核仁通过低离子强度缓冲液的匀浆和二价阳离子的螯合进行扩散。正常肝核仁共有的这些肿瘤核仁的抗原决定因子被兔抗肝核仁抗血清掩盖，形成免疫复合物。用这些抗体-核仁复合物免疫新西兰大白兔。所得抗血清在HeLa细胞中产生核仁荧光，但在正常人的肝脏或肾脏细胞中不产生。单、二维免疫印迹在HeLa细胞中鉴定出Mr 58000 /pl 5.8的主要抗原，而在正常人肝核仁的相应免疫印迹上未检测到该抗原。这种方法为肿瘤相关抗原的选择提供了一种改进的方法。
Nucleoli purified from HeLa cells were spread by homogenization in low ionic strength buffer and by chelation of divalent cations. The antigenic determinants of these tumor nucleoli that are shared by normal liver nucleoli were masked by addition of rabbit anti-liver nucleolar antisera to form immune complexes. New Zealand White rabbits were immunized with these antibody-nucleolar complexes. The resulting antisera produced nucleolar fluorescence in HeLa cells but not in normal human liver or human kidney cells. One- and two-dimensional immunoblots identified a major Mr 58,000/pl 5.8 antigen in HeLa cells, which was not detected on corresponding immunoblots of normal human liver nucleoli. This method offers an improved approach to selection of tumor-associated antigens.