Protein Biomarker Assays

Epic Sciences’ CTC Protein Biomarker analysis consists of:

1. Slide prep: Upon receipt of a patient blood sample, whole blood is lysed and nucleated cells are deposited on up to 12 microscope slides that are frozen at -80 ºC until analysis.
2. Cell staining and scanning: Slides are immunofluorescently stained and scanned by Epic Sciences’ rapid fluorescent scanning method, which images each nucleated cell.
3. CTC identification and biomarker analysis: A digital pathology algorithm, which includes protein expression and morphology, differentiates candidate CTCs from surrounding white blood cells (WBCs). CTCs are confirmed and classified into one of the following categories (1) Traditional CTCs, (2) CTC clusters, (3) CK(-) CTCs and (4) Apoptotic CTCs (see CTC basics to review these different classes of CTCs).

The open 4th and 5th channel can be integrated into our base assay for CTC identification and protein biomarker characterization. These are representative images (10x) of individual CLCs detected by our N-terminal Androgen Receptor (AR) assay. Blue = DAPI, red = Cytokeratin, green = CD45, white = N-terminal AR.

Biomarker expression data can be extracted and quantitated on each individual CTC. Below is an example of biomarker analysis that looks at androgen receptor (AR) expression on cell line cells. PC3s (AR negative) and LnCAPs (AR positive) were spiked into blood and analyzed by the Epic platform for AR expression. Each individual dot on the plot represents the AR expression of an individual CTC.

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