Underwood P A, Bean P A
CSIRO Division of Molecular Biology, North Ryde, NSW, Australia.
J Immunol Methods. 1988 Feb 24;107(1):119-28. doi: 10.1016/0022-1759(88)90017-8.
Three hybridoma clones, which were shown to change the characteristics of their antibody specificities when grown under different culturing conditions, are described in detail. This phenomenon was shown to be due to the persistence of mixed clones, even under conditions where standard statistical treatment indicated a high probability of monoclonality. Such mixed clones persisted, sometimes undetected, through repeated cycles of re-cloning. It was shown that the assumption that every viable clone has the same random chance of monoclonality, is invalid, and can lead to misleadingly high estimates for the probability of monoclonality. Verification of seeding of individual wells with single cells is recommended and the relative merits of this versus repeated limiting-dilution cloning are discussed.
详细描述了三个杂交瘤克隆，当在不同培养条件下生长时，它们的抗体特异性特征会发生变化。结果表明，即使在标准统计处理表明单克隆性概率很高的条件下，这种现象也是由于混合克隆的持续存在所致。通过重复的再克隆循环，这些混合克隆有时未被检测到而持续存在。结果表明，每个活克隆具有相同单克隆性随机机会的假设是无效的，并且可能导致对单克隆性概率的误导性高估。建议验证单细胞接种到各个孔中，并讨论了这种方法与重复有限稀释克隆的相对优点。
