CA125, is a marker in the clinical diagnosis of several cancers and currently is the best serum-based tumor marker for ovarian cancer. Here, we developed an ultrasensitive antibody-ssDNA aptamer sandwich-type fluorescence immunosensor for CA125 detection. Based on a novel signal amplification strategy the carbon dots (CDs) functionalized with aptamer (CD-aptamer) used as detection probe and PAMAM-Dendrimers/AuNPs was used for covalent attachment of CA125-antibody and completing the sandwich assay method. By measuring of fluorescence resonance energy transfer (FRET) signals between CDs and AuNPs as nanoquenchers, the fluorescence signal quenched during sandwich complex formed between anti-CA125, CA125 and CDs-Aptamer and decreasing of fluorescence response signal is related to CA125 concentrations. Under optimal conditions, the immunosensor exhibited an extremely low calculated detection limit of 0.5 f g/mL with wide linear range 1.0 fg/mL to 1.0 ng/mL of CA 125. The application of the immunosensor for CA125 detection in serum samples and measuring of ovarian-cancer cells was also investigated. The immunosensor revealed good sensitivity and specificity with ovarian cell concentrations from 2.5×103 to 2×104 cells/mL with correlation coefficient of 0.9937 and detection limit of 400 cells/mL (4 cell in 10 μL), indicating potential application of immunosensor in clinical monitoring of tumor biomarkers. Furthermore, the cell viability was not changed upon treatment with CDs probe during 24 h, showing the low cytotoxicity of the probe. More importantly, CDs-antibody hybrid was achieved in selective imaging of the cancer cells over the OVCAR-3 line cells, implying its potential applications in biosensing, as well as in cancer diagnosis.
