Sibley School of Mechanical and Aerospace Engineering, Cornell University, 245 Upson Hall, Ithaca, NY 14853, USA.
Biomed Microdevices. 2012 Apr;14(2):401-7. doi: 10.1007/s10544-011-9616-5.
Patients suffering from cancer can shed tumor cells into the bloodstream, leading to one of the most important mechanisms of metastasis. As such, the capture of these cells is of great interest. Circulating tumor cells are typically extracted from circulation through positive selection with the epithelial cell-adhesion molecule (EpCAM), leading to currently unknown biases when cells are undergoing epithelial-to-mesenchymal transition. For prostate cancer, prostate-specific membrane antigen (PSMA) presents a compelling target for immunocapture, as PSMA levels increase in higher-grade cancers and metastatic disease and are specific to the prostate epithelium. This study uses monoclonal antibodies J591 and J415-antibodies that are highly specific for intact extracellular domains of PSMA on live cells-in microfluidic devices for the capture of LNCaPs, a PSMA-expressing immortalized prostate cancer cell line, over a range of concentrations and shear stresses relevant to immunocapture. Our results show that J591 outperforms J415 and a mix of the two for prostate cancer capture, and that capture performance saturates following incubation with antibody concentrations of 10 micrograms per milliliter.
患有癌症的患者可能会将肿瘤细胞释放到血液中,这导致了转移的最重要机制之一。因此,这些细胞的捕获非常重要。循环肿瘤细胞通常通过上皮细胞黏附分子(EpCAM)的阳性选择从循环中提取,这导致在细胞经历上皮-间充质转化时出现目前未知的偏倚。对于前列腺癌,前列腺特异性膜抗原(PSMA)是免疫捕获的一个引人注目的靶标,因为 PSMA 水平在高级别癌症和转移性疾病中增加,并且特异性针对前列腺上皮。本研究使用针对活细胞中 PSMA 的完整细胞外结构域具有高度特异性的单克隆抗体 J591 和 J415-抗体,在与免疫捕获相关的一系列浓度和剪切应力下,在微流控设备中捕获表达 PSMA 的永生前列腺癌细胞系 LNCaPs。我们的结果表明,J591 优于 J415 和两者的混合物用于前列腺癌捕获,并且在与 10 微克/毫升的抗体浓度孵育后,捕获性能饱和。