Technology

Our Platform
BioFluidica's platform to analyze Circulating Tumor Cells, CTC's, consists of simple instrumentation that works in conjunction with a disposable, test-specific cartridge, approximately the size of a standard credit card. The test-specific cartridge can accept whole blood directly without any type of pre-processing which, simplifies CTC isolation, analysis, and improves results. After the operator loads the blood sample, all subsequent processing steps are performed by the associated instrumentation and cartridge. The processing steps include CTC capture, CTC washing and release, and CTC enumeration, using a highly sensitive, compact, and low-cost fluorescence microscope.

BioFluidica's CTC System
The heart of BioFluidica's CTC system, is a highly efficient CTC capture bed, comprised of 50 to 500 sinusoidally-shaped channels. These channels are coated with antibodies, which isolate specific CTCs from whole blood. The antibodies are chemically immobilized onto the surfaces of the capture bed using BioFluidicas patented technology. This allows for high recovery, over 95%, of CTCs from clinical samples and provides highly pure CTCs, over 80%, even when isolated from whole blood. Following release and enumeration, over 85% of the CTCs are viable. The cartridge is made from a plastic, which can be injection molded. Injection molding is the same technology used to produce plastic CDs and DVDs. As such, large numbers of cartridges can be produced at low-cost and with high replication fidelity, reducing assay cost and improving assay reproducibility.

Sinusoidal ChannelsFig.1 Biofluidica's patented CTC capture bed.

Efficiency
The isolated CTCs are released from the capture surface using a patented technology, which is simple to use and provides high release efficiency. Approximately 90% of the cells will be released in 30 minutes. This technology also provides a high rate of undamaged, viable cells, over 85%. The release strategy uses unique linkers that can be secured at low-cost and efficiently attached to the cartridge's surface in a very ordered fashion. These linkers can be clipped enzymatically to release the captured cells.

Reliability
Once the captured cells have been released, they can be swept through a highly specific, single-cell electrical sensor, where they are counted. This counting method eliminates the need for cell labeling and expensive optical detection. Because the cells are intact and viable and the purity of the CTCs is high, they can be used in downstream applications such as mRNA expression profiling, next generation sequencing and/or DNA mutational analysis.

Additional Features
Following release, BioFluidica’s instrumentation also can allow for (if required) the staining of CTCs and microscopic interrogation using the appropriate antibodies for immunophenotyping. The cartridge consists of units for the automated preparation of the cells for fluorescence interrogation and fitted with a highly sensitive fluorescence, yet compact and low-cost, microscope. Up to 4-colors can be interrogated using this microscope.

Tested & Proven
The CTC detection system has been tested in a variety of tumor types including breast, colorectal, ovarian, melanoma, pancreatic and prostate cancer. In pilot clinical studies where blood was collected from patients with metastatic disease and processed, the clinical sensitivity and specificity using BioFluidica's isolation technology has been demonstrated to be 100%. In addition, promising results have been obtained for acute myeloid leukemia and multiple myeloma when using the appropriate antibodies for capture.

Summary
Fully automated operation accepts whole blood samples directly.
High capture efficiency, over 95%, with high throughput, processes 10 mL of blood in less than 30 minutes, and with high purity, over 80%.
Disposable CTC cartridge made from a low-cost plastic in a high production mode.
Can release captured CTCs with high efficiency, over 90%, without damaging cells, giving a viability of over 85%
Label-less single cell counter using electrical signature of the single cells, coupled with the high purity of the cartridge, allows for CTCs to be subjected to a variety of molecular assays.
Low-cost instrumentation can fluorescently image the cells for immune-phenotyping if required

Technology