Circulating tumor cells: clinically relevant molecular access based on a novel CTC flow cell.
BACKGROUND
Contemporary cancer diagnostics are becoming increasing reliant upon sophisticated new molecular methods for analyzing genetic information. Limiting the scope of these new technologies is the lack of adequate solid tumor tissue samples. Patients may present with tumors that are not accessible to biopsy or adequate for longitudinal monitoring. One attractive alternate source is cancer cells in the peripheral blood. These rare circulating tumor cells (CTC) require enrichment and isolation before molecular analysis can be performed. Current CTC platforms lack either the throughput or reliability to use in a clinical setting or they provide CTC samples at purities that restrict molecular access by limiting the molecular tools available.


RESULTS
Recent advances in magetophoresis and microfluidics have been employed to produce an automated platform called LiquidBiopsy®. This platform uses high throughput sheath flow microfluidics for the positive selection of CTC populations. Furthermore the platform quantitatively isolates cells useful for molecular methods such as detection of mutations. CTC recovery was characterized and validated with an accuracy (<20% error) and a precision (CV<25%) down to at least 9 CTC/ml. Using anti-EpCAM antibodies as the capture agent, the platform recovers 78% of MCF7 cells within the linear range. Non specific recovery of background cells is independent of target cell density and averages 55 cells/mL. 10% purity can be achieved with as low as 6 CTCs/mL and better than 1% purity can be achieved with 1 CTC/mL.


CONCLUSIONS
The LiquidBiopsy platform is an automated validated platform that provides high throughput molecular access to the CTC population. It can be validated and integrated into the lab flow enabling CTC enumeration as well as recovery of consistently high purity samples for molecular analysis such as quantitative PCR and Next Generation Sequencing. This tool opens the way for clinically relevant genetic profiling of CTCs.
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Jessamine P
JP
Winer-Jones
Engineering, Cynvenio Biosystems, Westlake Village, California, United States of America.
Behrad
B
Vahidi
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Norma
N
Arquilevich
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Cong
C
Fang
Engineering, Cynvenio Biosystems, Westlake Village, California, United States of America.
Samuel
S
Ferguson
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Darren
D
Harkins
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Cory
C
Hill
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Erich
E
Klem
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Paul C
PC
Pagano
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Chrissy
C
Peasley
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Juan
J
Romero
Engineering, Cynvenio Biosystems, Westlake Village, California, United States of America.
Robert
R
Shartle
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Robert C
RC
Vasko
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
William M
WM
Strauss
Research and Development, Cynvenio Biosystems, Westlake Village, California, United States of America.
Paul W
PW
Dempsey
D000906:Antibodies / Q000737:chemistry; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000235:genetics; D057205:Automation, Laboratory; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics; D002452:Cell Count; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D000071858:Epithelial Cell Adhesion Molecule; D015870:Gene Expression; D006801:Humans; D059346:Magnets; D046210:Microfluidic Analytical Techniques; D009360:Neoplastic Cells, Circulating / Q000378:metabolism* / Q000473:pathology; D012212:Rheology
D000906:Antibodies; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule