Sensitive on-chip detection of cancer antigen 125 using a DNA aptamer/carbon nanotube network platform

Despite the major role of cancer antigen 125 (CA125) in cancer progression, its structural diversity makes antibody-based immunoassay of this protein difficult. Here we report the selection of an anti-CA125 ssDNA aptamer and its application to sensitive detection of CA125 when used together with a CA125-specific antibody immobilized on a three-dimensional network of carbon nanotubes (3DN-CNTs). With exceptional stability and easy modifications over antibodies, the selected ssDNA aptamer (rCAA-8) showed high binding affinity (166 nM) for recombinant CA125, which enabled CA125-specific imaging in ovarian cancer cells (OVCAR-3). Furthermore, when compared to other fluorescent assays based on graphene oxide or flat surface and traditional enzyme-linked immunosorbent assay, a chip-based assay using a 3DN-CNT surface and anti-CA125 antibody-aptamer pair resulted in higher sensitivity and broader dynamic range as a function of CA125 concentration, due to high target specificity and high surface loading density. Our on-chip aptamer-based assay will facilitate sensitive and specific monitoring of CA125 in the biological and clinical fields.