Collaborations with External Organisations and Service Providers
This equipment currently provides the main focus of CTC enumeration for trial and experimental medicine projects within the CTC team. Approved by the US FDA for CTC enumeration in key disease areas including lung cancer, the system “captures” CTCs from patient blood samples using an antibody to Epithelial Cell Adhesion Molecule (EpCAM) coupled to microscopic ferrous beads. Following capture and staining for DNA, cytokeratins and CD45, the CTC images are viewed under a fluorescent microscope and counted by a trained analyst.
Due to the marker-dependent capture approach, this system is most suited to epithelial cancers such as SCLC. Janssen, the manufacturers of the CellSearch, are currently developing a new marker-independent system and CEP are discussing with Janssen the possibility of trialling this in our laboratories.
RareCyte provide a novel technology platform which, without prior enrichment of CTCs or depletion of leukocytes, enumerates and isolates CTCs from blood. The platform includes the AccuCyte® blood separation system and CyteSealer® which allow precise separation of nucleated cells and plasma. The platform also includes CyteSpreader® slide preparation device, CyteFinder® instrument (for cell imaging with analysis) and the CytePicker® for retrieving automatically pre- identified rare cells for further single-cell downstream analysis including genomic or proteomic analysis. The system is demonstrated to have high recovery in spiked blood samples (90-95%) and CEP will be evaluating it in Non- Epithelial tumour types such as Non-Small Cell Lung Cancer (NSCLC) and Melanoma.
Clearbridge ClearCell FX1
The ClearCell FX1®, provided by Clearbridge Biomedics, is a microfluidic biochip based platform capable of enriching CTCs from blood based on size, deformability and inertia via inherent Dean Flow Fractionation in spiral channels. Enrichment takes place from 7.5ml of blood in less than an hour and appropriately collected cells are viable at the end of the enrichment process. Enriched Cells are suitable for downstream analysis.
Aushon Biosystems provide the multiplex ELISA capability in CEP for the analysis of patient plasma and serum samples for circulating biomarkers. Aushon have upgraded their system from the previous “square-plex” plate format to the circular “CiraPlex” format, and this comes with improved assay sensitivity and plate stability. The CEP project portfolio currently includes a number of active and planned projects requiring ELISA analysis for a range of circulating biomarkers, including angiogenesis factors and cell death biomarkers, and this is set to continue. Therefore, continued development and refinement of the multiplex ELISA capability is an important feature of the GCP Team’s activity going forward.
The CellCelector™ combines precise robotic technology (positional accuracy +/-1μm range) with an imaging software package and an embedded inverted microscope to provide live imaging, specific selection and a fully documented picking process by using brightfield as well as fluorescence illumination. Included is a software-controlled filter for up to six fluorescence excitation filters to enable the detection of various fluorochromes.
The CellCelector™ technology represents a powerful tool for the isolation of single cells or colonies. This will complement the DEPArray technology for single CTC isolation, and also allows the possibility of isolating groups of CTCs (Circulating Tumour Microemboli, or CTM), and also live cells giving the potential for short-term culture.
GILUPI® CellCollector (Detektor Cancer01 DC01) device positively enriches CTCs in vivo based on EpCAM expression via EpCAM-coated gold or platinum wires. The process involves insertion of the wire into a patient’s vein for 30min, which achieves an estimated blood volume exposure 20ml per minute. Detection and characterisation of CTCs thus captured are carried out with the cells still bound to the wire.