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Rushton, A. Circulating Tumour Cell Enrichment Technologies. Encyclopedia. Available online: https://encyclopedia.pub/entry/7974 (accessed on 19 December 2025).
Rushton A. Circulating Tumour Cell Enrichment Technologies. Encyclopedia. Available at: https://encyclopedia.pub/entry/7974. Accessed December 19, 2025.
Rushton, Amelia. "Circulating Tumour Cell Enrichment Technologies" Encyclopedia, https://encyclopedia.pub/entry/7974 (accessed December 19, 2025).
Rushton, A. (2021, March 12). Circulating Tumour Cell Enrichment Technologies. In Encyclopedia. https://encyclopedia.pub/entry/7974
Rushton, Amelia. "Circulating Tumour Cell Enrichment Technologies." Encyclopedia. Web. 12 March, 2021.
Circulating Tumour Cell Enrichment Technologies
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This entry is adapted from the peer-reviewed paper 10.3390/cancers13050970

Circulating tumour cells (CTCs) are the precursor cells for the formation of metastatic disease.

circulating tumour cell (CTC) cancer metastasis liquid biopsy

1. Background

With a simple blood draw, liquid biopsies enable the non-invasive sampling of CTCs from the blood, which have the potential to provide important insights into cancer detection and monitoring. Since gaining FDA approval in 2004, the CellSearch system has been used to determine the prognosis of patients with metastatic breast, prostate and colorectal cancers. This utilises the cell surface marker Epithelial Cell Adhesion Molecule (EpCAM), to enrich CTCs, and many other technologies have adopted this approach. More recently, the role of mesenchymal-like CTCs in metastasis formation has come to light. It has been suggested that these cells are more aggressive metastatic precursors than their epithelial counterparts; however, mesenchymal CTCs remain undetected by EpCAM-based enrichment methods. This has prompted the development of a variety of ‘label free’ enrichment technologies, which exploit the unique physical properties of CTCs (such as size and deformability) compared to other blood components.

2. Introduction

Circulating tumour cells (CTCs) are shed into the bloodstream from both primary and metastatic tumours and those that are able to survive in the circulation represent metastatic precursor cells [1]. CTCs are important biomarkers for disease and are a powerful tool to study tumour progression and evolution. They represent a rare and heterogeneous population of cells, typically accounting for ∼1 cells for every 105–106 peripheral blood mononuclear cells (PBMCs), so a key challenge for their clinical utility is the development of standardised isolation and characterisation technologies [2]. There are numerous technologies that have been developed to enrich CTCs from normal hematopoietic cells that rely on physical and biological properties of CTCs, including size, density, cellular charge and expression of cellular markers. The enrichment techniques (Table 1) can broadly be divided into immunocapture methods that differentiate cells based on epithelial cell surface marker expression, notably epithelial cell adhesion molecule (EpCAM) (Figure 1A), and those that differentiate based on distinct biophysical properties (Figure 1B,C). If CTC enrichment and characterisation is to be routinely used in the clinical setting, technologies must ideally meet several criteria: they must have high detection and recovery rates, with accurate throughput sample processing and enumeration capability. Further, they must be generally fully automated and easy to use, with little to no pre-processing of blood required. Finally, if they are to have wide clinical applicability, they must be able to detect heterogeneous cells from a wide range of different cancers.

Cancers 13 00970 g001 550
Figure 1. Summary of circulating tumour cell (CTC) enrichment technologies. (A) Immunocapture methods including immunomagnetic positive and negative enrichment methods, microfluidic immunocapture methods, nanomaterial immunocapture enhancement and their relevant technologies; (B) Biophysical property enrichment methods including membrane filtration, size-based microfluidics, density based and dielectrophoresis and associated technologies; (C) Other methods including in vitro, combined and secondary isolation methods and associated technologies.

Table 1. CTC isolation technologies, grouped based on enrichment method. Capture efficiency, recovery rate and advantages and disadvantages of the technologies are also shown.

Subcategory Name Capture Efficiency (%) Recovery Rate (%) Advantages Disadvantages
Immunomagnetic enrichment          
Immunomagnetic positive enrichment CellSearch [3][4][5][6][7] 42–90   Semi automated
Can process up to 8 samples at a time
In device staining
CTC enumeration via CellTracks Analyser
FDA approved		 Recovery of EpCAM+ CTCs only
Only able to detect CTCs expressing high levels of EpCAM
  MACS [8][9] 25–90   Cocktail of antibodies available to increase CTC capture
Able to process up to 15 mL blood
Easy elution of CTCs
Pro Separator can process up to 6 samples at once		 Recovery of EpCAM+ CTCs only
Suggested the MACS system is better suited for tissue samples
  MagSweeper [10][11] 60–70   Nonadherent plastic sleeves allow for multiple rounds of capture to increase capture efficiency Recovery of EpCAM+ CTCs only
  Strep-tag [12][13] 79–86 70 Easy release of CTCs by simple addition of d-biotin
Possibility to use a cocktail of antibodies to increase capture		 Recovery of EpCAM+ CTCs only
  IMS [14] 92   Leukocytes repelled so high purity recoveries Recovery of EpCAM+ CTCs only
Not yet tested on patient samples
Immunomagnetic negative enrichment EasySep [15][16] 19–65   Recovery of heterogeneous population of CTCs Exclusion of CTC-WBC clusters
Variable recovery rates
May inadvertantly remove CTCs
  RosetteSep [17] 62.5   Recovery of heterogeneous population of CTCs
Cocktail of antibodies used to maximise depletion		 Exclusion of CTC-WBC clusters,
May inadvertantly remove CTCs
Microfluidic immunocapture positive enrichment CTC-Chip [18][19]   >60 Large surface area for CTC capture
High viability of recovered cells		 Recovery of EpCAM+ CTCs only
Slow processing rate
Complex geometry of chip difficult to scale up
Geometry prevents passage of CTC clusters
  HB-chip [20] 74.5–97   HB grooves increase CTC-antibody contact for increased cell capture Recovery of EpCAM+ CTCs only
  GEDI chip [21] 80–90   Large surface area for CTC capture
Possibility to functionalise with alternative antibodies		 May miss heterogeneity of CTCs
  HTMSU [22] >97   Quick processing
On-chip single-cell conductometric counting for enumeration		 Recovery of EpCAM+ CTCs only
  Nanovelcro [23] 70–95   4 generations developed for different clinical utilities
3rd and 4th generation chips adapted for easy CTC release		 Recovery of EpCAM+ CTCs only
  Isoflux [4] 74–90 64–75 Utilises microfluidic approach to increase EpCAM sensitivity
Up to 4 samples can be processed in parallel
Multiple kits including cocktails of antibodies to capture heterogeneity
IsoFlux Cytation Imager for sample scanning		  
Capture enhancement by nanomaterials NP-HBCTC-Chip [24] 79–97   Simple release of CTCs by addition of glutathione (GSH)
Chip surface can be functionalised with a cocktail of antibodies for enhanced capture efficiency		 Recovery of EpCAM+ CTCs only
Very low throughput
  GO chip [25][26] 67–100 91–95 Simple chip design
Large surface area for increased CTC capture		 Recovery of EpCAM+ CTCs only
  SiNP [27] 84–91   Large surface area for CTC capture Recovery of EpCAM+ CTCs only
Capture enhancement by nanomaterials Nanotube-CTC-chip [28] 89–100   Preferential adherence negates need for EpCAM antibodies
Planar enrichment surface makes chip visualisation and imaging easy		 Time taken for optimal CTC adherence to substrate is too long
Size based enrichment          
Membrane filtration FMSA [29] 90   Recovery of heterogeneous population of CTCs
Cheap and easy to produce
Quick processing time		 Filter clogging highly likely
  ScreenCell [30]   74–91 Recovery of heterogeneous population of CTCs
Cheap and easy to produce
Three different devices offered depending on downstream requirements
Quick processing time		 Unevenly distributed or fused pores can reduce capture efficiency
  ISET [31][32]   83–100 Recovery of heterogeneous population of CTCs
Cheap and easy to produce
Ability to process 12 samples in parallel		 Slow processing time
Blood must be diluted 1:10 to prevent membrane clogging
  SB microfilter [33] 78–83   Recovery of heterogeneous population of CTCs
Cheap and easy to produce
Quick processing time		 Only 1 mL blood can be processed at a time due to device clogging
  FAST [34]   94–98 Recovery of heterogeneous population of CTCs
Cheap and easy to produce
Quick processing time		  
Microfluidics Parsortix [35] 42–70 54–69 Recovery of heterogeneous population of CTCs
Ability to capture CTC clusters
Option for on-chip staining		 Slow processing time
On-chip imaging difficult
  MCA [36] >90 68–100 Recovery of heterogeneous population of CTCs
Option for on-chip staining
Ability to process up to 4 samples in parallel		  
  ClearCell FX1 [37][38]   52–79 Recovery of heterogeneous population of CTCs
Quick processing time
No channel clogging observed		  
  Vortex VTX-1 [39][40]   53.8–71.6 Recovery of heterogeneous population of CTCs
Filters at channel inlet prevent channel clogging
Fully automated process
Quick processing time
Associated BioView for enumeration
Option to run in “high recovery” or “high purity” mode		  
  p-MOFF [41]   91.6–93.75 Recovery of heterogeneous population of CTCs
Quick processing time
No channel clogging observed		 RBC lysis and Ficoll density centrifugation required
Density based OncoQuick [42][43] 25–87   Recovery of heterogeneous population of CTCs
Up to 25 mL blood can be processed per tube		 Low detection and recoveryrates
  AccuCyte [44]   81–90.5 Recovery of heterogeneous population of CTCs
Allows for processing of multiple samples in parallel
Associated CyteFinder and CytePicker systems for imaging and mechanical selection of CTCs		  
Other          
Dielectrophoresis ApoStream [45][46]   55–78.5 Recovery of heterogeneous population of CTCs
Quick processing time
iCys laser scanning cytometer for enumeration
High viability of recovered cells		  
In vivo Diagnostic leukapheresis (DLA) [47]     Recovery of heterogeneous population of CTCs
Recovery of much greater numbers of CTCs		 Only a pre-enrichment step so must be used in combination with another enrichment technology
Huge leukocyte background
  GILUPI CellCollector [48]     Potential for much greater numbers recovered More invasive for the patient than a simple blood draw
Recovery of EpCAM+ CTCs only
Combined CTC-iChip [49]   70–100 Option for positive or negative enrichment approach
Inertial focusing provides high sensitivity selection
Quick processing time		 Positive enrichment only allows for recovery of EpCAM+ CTCs
Negative enrichment will exclude CTC-WBC clusters
  LPCTC-iChip [50]   85.5–100 Potential for much greater numbers recovered
Magnetic field directs WBCs to centre of channel to prevent channel clogging
Extremely high throughput		 Disregards CTC-WBC clusters
Initial debulking step may result in CTC loss
  OPENchip [51]   50 Chip allows for CTC enrichment and on-chip downstream molecular analysis Low throughput, low recovery rates

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