Detection of circulating prostate cancer cells by means of an antibody-conjugated nanoparticular biosensor

M. Raschid Hoda, Gerit Theil, Ekkehard Weber, Klaus Lücke, Paolo Fornara
Clinic for Urology and Kidney Transplantation Centre
University Medical School of Halle/Wittenberg, Germany


Introduction:
Currently existing techniques for the isolation of circulating tumor cells in blood (CTC) have experimental approaches. Nanoparticles as biosensors could allow a refinement of the method and possible clinical applicability of the CTC-isolation in cancer patients.

Methods:
In a translational project will be tested at the clinic, the clinical application of a coated with anti-EpCAM antibody-nano-scaled detector for the isolation of CTCs from peripheral blood of patients with prostate cancer. We reported on the results of preclinical testing.

Results:
The Nanodetektor consists of a nano-gold particles loaded with steel wire (GILUPI GmbH, Potsdam). This was coated with anti-EpCAM-AK. At first, the characterization of EpCAM expression in established PCa cell lines DU 145, PC3 and LNCaP cells as a model for various tumor entities. Cytochemistry showed that all three established cell lines express EpCAM on the cell membrane. The next step was to determine the cell binding efficiency of an anti-EpCAM-functionalized Nanodetektors in the fluid-dynamic system (Figure 1). The cell-binding experiments with cell lines showed a sufficient covalent binding of cells to the detector. For the characterization of the bound CTC was the multiplex RT-PCR (AdnaTestProstateCancerDetect) is used, the tumor antigens, PSMA, PSA, and EGFR were detected. In another step, blood samples from 20 prostate cancer patients of different stages were studied. These blood samples were used in the fluid dynamic system (Fig. 2). The characterization of isolated cells was carried out on the RT-PCR with regard to the marker PSMA, PSA, EGFR and cytochemical level on the detection of EpCAM expression and the control CD45-staining (Fig. 3). These were isolated on average 42.92 and 3.41 CTC/7.5ml blood at hrPCA CTC/7.5ml in locally limited PCA.

Conclusion:
Isolation of CTC using an antibody-coated nano-detector from the blood of prostate cancer patients of different stages is possible with high efficiency. The first in-vivo use of this system is currently being tested clinically.

 
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