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© 2016 ANGLE Europe Ltd.. All rights reserved. Parsortix™: liquid biopsy for companion diagnostics
© 2016 ANGLE Europe Ltd.. All rights reserved.
Parsortix™: liquid biopsy for companion diagnostics
© 2016 ANGLE Europe Ltd.. All rights reserved. 2
Circulating tumor cells and companion diagnostics
CTCs are central to metastases formation
© 2016 ANGLE Europe Ltd.. All rights reserved. 3
Parsortix CTC enrichment technology
Stepped, microscale cell separators for fluid flow and cell separation
Two granted US Patents
Granted patents in China, Canada and Australia
Patents pending worldwide
European patent granted
© 2016 ANGLE Europe Ltd.. All rights reserved. 4
Parsortix cell capture
Inlet
Outlet
Patented Separation Step
Plan View
Cross Section
Captured CTCs
White blood cells Red blood cells pass through
Blood Flow
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Cells stained fluorescently in the cassette
HT29 cell captured and stained in-cassette using CK20 (green), DAPI (blue). Source: University of Surrey (Annels/Pandha).
HT29 cells captured and stained in-cassette using CK20 (green), DAPI (blue). Note: brightfield on allowing visualisation of the ‘steps’ in the cassette. Source: University of Surrey (Annels/Pandha).
CTC captured and stained in-cassette from colo-rectal cancer patient sample. CK20 (green), TO-PRO 3 nuclear stain (blue). Source: University of Surrey (Annels/Pandha).
© 2016 ANGLE Europe Ltd.. All rights reserved. 6
Cells harvested for identification and analysis
Breast cancer cell line, captured, harvested and stained. CK (orange); CD45 (red) DAPI (blue). Source: Universitätsklinikums Hamburg-Eppendorf (Gorges/Pantel).
CTC harvested and stained from colorectal cancer patient sample. CK (orange); DAPI (blue). Source: Universitätsklinikums Hamburg-Eppendorf (Gorges/Pantel).
CTCs harvested and stained from breast cancer patient sample. CK (orange); DAPI (blue). Source: Universitätsklinikums Hamburg-Eppendorf (Gorges/Pantel).
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Range of analytical downstream workflows
• Immuno-fluorescence (in cassette or on harvest)
• Gene expression analysis • DNA mutation analysis • FISH
• Feed DEPArray/CellCelector followed by single cell WGA and NGS
• Gene expression analysis • DNA mutation analysis
cfDNA removed prior to capture of CTCs
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…….used by leading cancer research groups in Europe and North America
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Key findings:
• Parsortix enables processing a single blood sample for cfDNA and CTCs
• For EpCAM expressing cells – no difference between CellSearch and Parsortix
• Cell lines with poor EpCAM expression – only Parsortix recovers cells
• CK+ve CTCs found in 12/12 SCLC patients (compared to 10/12 for CellSearch)
• Cells imaged and counted on DepArray
Ged Brady, Cancer Research UK Manchester Institute “The Parsortix system has a unique combination of features making it suitable for routine clinical analysis of patient blood samples. We have now incorporated the Parsortix workflow into multiple clinical trials and have been accumulating many hundreds of stored enriched samples that will be of immense value in our future CTC studies.”
“Clinical evaluation of a novel microfluidic device for epitope-independent enrichment of circulating tumour cells in patients with small cell lung cancer”, Analyst Dec 2015
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Key findings:
• Easy accessibility to viable CTCs in liquid suspension.
• Parsortix harvests intact and viable tumour cells. Cells still functional for further molecular characterisation
• mRNA expression of specific transcripts on harvested cells and array CGH illustrates suitability for molecular analysis .
• High purity of harvest with background WBC in 1000-2000 range
“A novel microfluidic platform for size and deformability based separation and the subsequent molecular characterisation of viable circulating tumor cells”, IJC, Jan2016
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Key findings:
• Use of a ficoll gradient as a pre-preparation on the blood sample allows processing of larger volumes of blood
• In prostate cancer patient samples, Parsortix harvested more CTCs than antibody based systems
• Purity of cell harvest from Parsortix was well ahead of antibody based systems and suitable for FISH analysis.
• Cells with mesenchymal markers can be detected on cells captured by Parsortix
• Cells harvested by Parsortix are viable and can be cultured.
“Optimization and Evaluation of a Novel Size Based Circulating Tumor Cell Isolation System ”, PlosOne, Sept 2015
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Key findings:
• Demonstrated use of gene expression analysis on the harvest from Parsortix
• High detection of CTC related transcripts even in early stage patients (ovarian cancer)
• qPCR based analysis of CTCs in harvest provides high throughput analysis and user independency
Development of clinical application:
• Gene expression panel for reliable detection of ovarian cancer in early stage patients has shown great promise
• Currently building larger data set to establish sensitivity/ specificity
• Goal is a ‘triaging’ assay to help determine risk of malignancy prior to first surgical procedure
“Circulating rare cells enable highly efficient cancer detection”, Poster, AACR 2015
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Summary
Why use Parsortix for CTC liquid biopsy?
• Parsortix captures cells that aren’t being caught by antibody based systems
• Easy access to the CTCs to feed a wide range of downstream analysis
• Possible to do ctDNA and CTCs from the same sample
• Cells are viable
• Used by leading research groups/ CE marked (IVDD)
• Simple, flexible, validated system
