In lung cancer, the AdnaTest CTC-Select provides antibodies against EpCam, Her2, EGFR, resulting in a strong increase of performance compared to detection based only on EpCam (Hanssen et al. (2016) Nature Sci Rep. 6, 28010).
In the first step, the CTCs in the blood are enriched (AdnaTest Select). This is achieved using antibody-coated magnetic particles (beads). Several antibodies are used, which bind with high specificity and affinity to the corresponding cancer cells. The enriched cells are lysed and subsequently purified several times to extract mRNA.
This step consists of mRNA purification, reverse transcription and subsequent analysis of tumor associate markers by multiplex PCR. Resulting fragments are analyzed on a Bioanalyzer 2100.
The first lane shows the DNA size standard (DNA-Ladder). Samples 1 is positive for ALDH1 and GA733.2, sample 3 is positive for ALDH1 and sample 2 is negative. The PCR negative control and the positive control (LungDetect) are shown in the last two lanes.
The first lane shows the DNA size standard (DNA-Ladder). Sample 1 is negative. Sample 2 is positive for TWIST1, sample 3 is positive for Akt-2 and PI3Kα, and sample 4 is positive for PI3Kα and TWIST1. Signals for all three EMT-2 markers can be detected in sample 5 and actin is detected in samples 1 to 5. The PCR negative control (C-) and the positive control (EMT-2) are shown in the last two lanes.