A Hemin-Graphene Nanocomposite-Based Aptasensor for Ultrasensitive Colorimetric Quantification of Leukaemia Cells Using Magnetic Enrichment

Diagnostic blood cell counting is of limited use in monitoring a minimal number of leukaemia cells, warranting further research to develop more sensitive and reliable techniques to identify leukaemia cells in circulation. In this work, a hemin-graphene nanocomposite-based aptasensor was developed for ultrasensitive colorimetric detection of leukaemia cells (CEM) using magnetic enrichment. Hemin-conjugated graphene oxide nanocomposites (HGNs) were prepared by hydrazine reduction using graphene oxide nanosheets and hemins. Hence, the prepared HGNs become able to absorb single-stranded DNA and acquire peroxidase-like activity. The aptamer sgc8c, which recognizes a specific target on leukaemia cells, was absorbed onto HGNs to capture the target CEM cancer cells. The captured target cells that associated with the HGNs were then concentrated and separated by magnetic beads (MBs) coated with sgc8c aptamers, forming a HGN-cell-MB sandwich structure. These sandwich structures can be quantified via an oxidation reaction catalysed by HGNs. By utilizing dual signal amplification effects generated by magnetic enrichment and the improved peroxidase activity of HGNs, the biosensor allowed for highly sensitive detection of 10 to 10(5) CEM cells with an ultra-low limit of detection (LOD) of 10 cells under optimal conditions. It is expected that the proposed aptasensor can be further employed in monitoring the minimal residual disease during the treatment of leukaemia.