Abstract

Getting Single Cells out of High Volume Liquid Biopsy

Klaus Drese, Director, "Coburg University of Applied Scienence and Arts "

Future cancer therapy is expected to rely heavily on the so called “liquid biopsy”. Tumor cells which circulate in the blood of cancer patients (CTCs) will be used to gain detailed diagnostic information on tumor subtypes to tailor the therapeutic intervention for each patient (“personalized medicine”). CTCs are extremely rare and it is commonly understood, that already a few CTCs in 7.5 mL of blood have diagnostic relevance. Even enumerating the CTCs is a challenge. Here, we report on a fully automated process to isolate the CTCs starting out directly from a 7.5 mL blood vial. The process combines large volume liquid handling for pre-enrichment of the CTCs with a microfluidic cartridge. The pre-enrichment is based on immuno-magnetic methods, removes most of the other blood constituents and reduces the volume from 7.5 mL to 500 µL. The pre-enriched CTCs are then automatically transferred to a microfluidic cartridge. On the cartridge, the CTCs are detected by a miniaturized flow cytometry module and each cell is individually dispensed into a well of a standard micro-titer plate. The overall process takes around one to two hours and delivers isolated single CTCs ready to be analyzed by methods such as next generation sequencing.