Homogeneous immunoassay for alpha-fetoprotein based on the quenching of the fluorescence of quantum dots by antibody labelled with complexed copper ion tags.

Abstract

A homogeneous fluorescent immunoassay is described for the determination of alpha fetoprotein (AFP) relying on the interaction between copper ion complex and quantum dots (QDs). Thecopper ion complex-labelled antibody can be employed as a quencher of fluorescence of QDs and capture probe of AFP in homogeneous solution. The labelled antibody is mixed with QDs to form the immune ensemble probe. Upon the addition of AFP, the labelled antibody is stripped away from QDs by antigen-antibody combination leading to the increase in the fluorescence signal. Thus, the determination of AFP can be realized by fluorometry (best measured at excitation/emission wavelengths of 360/520nm). The fluorescence intensity shows a good linear relationship with the AFP concentration ranging from 40 to 640ngmL-1, and the LOD is 26ngmL-1. The proposed method provides a new approach to incorporate metal complexes into QD-based biomolecule sensing. Graphical abstract Schematic presentation of a fluorescent probe comprised of quantum dots and antibody labelled with copper ion complex for homogeneous immunoassay of α-fetoprotein. The target antigen can break up the ground state QD/labelled antibody complex to set free the fluorescent QDs.