Enrichment of rare circulating tumor cells (CTCs) in blood is typically achieved using antibodies to epithelial cell adhesion molecule (EpCAM), with detection using cytokeratin (CK) antibodies. However, EpCAM and CK are not expressed in some tumors and can be downregulated during epithelial-to-mesenchymal transition. A micro-fluidic system, not limited to EpCAM or CK, was developed to use multiple antibodies for capture followed by detection using CEE-Enhanced (CE), a novel in situ staining method that fluorescently labels the capture antibodies bound to CTCs. Higher recovery of CTCs was demonstrated using antibody mixtures compared to anti-EpCAM. In addition, CK-positive breast cancer cells were found in 15 of 24 samples (63%; range 1–60 CTCs), while all samples contained additional CE-positive cells (range 1–41; median = 11; P = .02). Thus, antibody mixtures against a range of cell surface antigens enables capture of more CTCs than anti-EpCAM alone and CE staining enables the detection of CK-negative CTCs.
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机译：通常使用抗上皮细胞粘附分子（EpCAM）的抗体实现血液中稀有循环肿瘤细胞（CTC）的富集，并使用细胞角蛋白（CK）抗体进行检测。但是，EpCAM和CK在某些肿瘤中不表达，在上皮到间充质转化过程中可能被下调。开发了一种不限于EpCAM或CK的微流体系统，以使用多种抗体进行捕获，然后使用CEE-Enhanced（CE）进行检测，这是一种新颖的原位染色方法，可以荧光标记与CTC结合的捕获抗体。与抗EpCAM相比，使用抗体混合物证明了CTC的更高回收率。此外，在24个样本中的15个样本中发现CK阳性乳腺癌细胞（63％；范围1–60 CTC），而所有样本均包含其他CE阳性细胞（范围1–41；中位数= 11； P = .02 ）。因此，与单独的抗EpCAM相比，针对一系列细胞表面抗原的抗体混合物能够捕获更多的CTC，而CE染色能够检测CK阴性的CTC。
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