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“Isolation of rare circulating tumour cells in cancer patients by microchip technology” Nagrath et al. Nature 2007 Peter Bojo Big Picture • Potential uses of CTCs – Choice of therapy – Monitoring of therapy – Explaining the biology of metastasis • Microchip is one of many CTC technologies – Immunomagnetic beads – Total RNA qPCR Microchip Design • • • • Blood pumped through forest of posts ~80,000 posts on a ~10cmX10cm chip EpCAM antibodies on posts Testing with cancer cell spiked solutions determines: – Flow rate ~1mL/hr – High capture efficiency even for low EpCAM expression levels ~60% – Identical capture efficiency for whole blood and lysed blood Microchip Workflow 1. Take whole blood sample, flow through optimized chip 2. Stain with DAPI, cytokeratin, and CD45 antibodies. 47% purity 3. Analyze CTC sensitivity, viability, and predictive ability Cancer Patient CTC Experiment • NSCLC, prostate, pancreatic, breast, and colon cancer samples. • Metastatic and non-metastatic prostate cancer samples. • Isolated between 5 and 1300 CTCs per patient in 115/116 cases. • 0 CTCs in healthy samples • High sensitivity Predictive Ability for Cancer Treatment Conclusions • Microchip is a competitive CTC enrichment technology • Uses unprocessed blood • High purity of isolated cells: 47% • High sensitivity: 99% • Capture efficiency: ~60% • High cell viability: 98% • Further Study warranted for: – CTC count and tumor size correlation – Comparable numbers of CTCs in localized and metastatic cancers