A DNA dendrimer amplified electrochemical immunosensing method for highly sensitive detection of prostate specific antigen

Abstract

This work designed a DNA dendrimer for the loading of signal molecule and the construction of amplified electrochemical immunosensing method. The DNA dendrimer was self-assembled by the hybridization of one couple of complementary oligonucleotides (DNA and cDNA) that were covalently conjugated to three arms of a Y-shaped cross-linker, tris(2-maleimidoethyl)amine (TMEA) respectively. The immunosensor was prepared by coating chitosan on glassy carbon electrode to covalently immobilize the capture antibody with glutaraldehyde as a linker. After the target protein was captured on the immunosensor, cDNA-labeled secondary antibody was bound on the surface via a sandwiched immunoreaction to introduce the DNA dendrimer onto immunosensor for loading abundant methylene blue as signal molecule, which amplified greatly the amperometric signal for immunoassay. Using prostate specific antigen (PSA) as a model analyte, this proposed method showed a wide linear range from 1 pg mL−1 to 10 ng mL−1 along with a limit of detection down to 0.26 pg mL−1. The designed strategy avoided complex synthesis of signal tags, and possessed excellent performance for analysis of practical samples, thus providing a new avenue for the development of signal amplification strategy and immunoassay methods.