Multiplex electrochemiluminescence immunoassay of two tumor markers using multicolor quantum dots as labels and graphene asconductingbridge

A multiplex electrochemiluminescence (ECL) immunoassay for simultaneous determination of two different tumor markers, alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA), using multicolor quantum dots as labels and graphene as conducting bridge was developed. Herein, a typical sandwich immune complex was constructed on the glass carbon electrode, with QDs525 and QDs625 labeled on secondary anti-AFP and anti-CEA antibodies, respectively, thus to obtain distinguishable ECL signals. Because most of those QDs labeled on secondary antibodies were beyond the space domain of the ECL reaction, graphene was used as a conducting bridge to promote the electron transfer between QDs and the electrode, leading to about 30-fold enhancement of the ECL intensity. Experimental results revealed that the multiplex electrochemiluminescence immunoassay enabled the simultaneous monitoring of AFP and CEA in a single run with a working range of 0.001–0.1 pg/mL. The detection limit (LOD) for both analytes at 0.4 fg/mL was very low. No obvious cross-reactivity was found. Precision, recovery and stability were satisfactory. This novel multiplex ECL immunoassay provided a simple, sensitive, specific and reliable alternative for the simultaneous detection of tumor markers in clinical laboratory.

► A multiplex ECL immunoassay of two tumor markers was proposed.
► Multicolor QDs were labeled to obtain distinguishable ECL signals.
► G was used as a conducting bridge, improving the ECL intensity about 30-folds.
► LOD for AFP and CEA was about 0.4 fg/mL.