| Summary: | 碩士 === 國立中山大學 === 醫學科技研究所 === 107 === We have developed self-linkable magnetic graphene oxide-bPEI-prussian blue (MPP) as a peroxidase mimicking nanozyme with high oxidizability to 3,3’,5,5’-tetramethylbenzidine (TMB), which generates significant absorption intensity for the colorimetric immunosensing of apolipoprotein A1 (ApoA1) in early bladder cancer (BC) diagnosis and prognosis monitoring. The ultrasensitive immunosensor was constructed using an ApoA1 antibody (ApoA1Ab)-functionalized chip (biochipApoA1) and self-linkable magnetic graphene oxide-bPEI-prussian blue (MPP). After incubating the sample and capturing ApoA1 proteins captured on the biochipApoA1, the MPP functionalized with ApoA1Ab and mouse IgG (MPP-1), rabbit anti-mouse IgG antibody (MPP-2), and goat anti-rabbit IgG antibody (MPP-3) were added together. We envisioned that each captured ApoA1 protein would allow the retention of a large amount of MPP through a self-linking process to amplify the colorimetric signal of TMB in the presence of H2O2. The linear detection range could be obviously widened in the presence of self-linkable MPP—from 0.05 ng/mL to 100 ng/mL—compared with the group without signal amplification (from 1 ng/mL to 100 ng/mL). Our immunosensor analysis of ApoA1 in the urine of BC patients and healthy individuals was highly correlated with enzyme-linked immunosorbent assay measurements; moreover, the ApoA1 concentrations of patients with high-grade BC were significantly higher than those of patients with low-grade BC. After standard clinical treatment, a significant drop of ApoA1 concentration occurred in urine that was lower than the cut-off concentration, suggesting potential clinical applications of the new self-linkable MPP-generating colorimetric immunosensor in early BC diagnosis and prognosis monitoring.
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