Development of sensitive
homogeneous assays is a high-priority
research target for clinical diagnostics. Quantum dots (QDs) present
favorable photophysical properties, which implies their potential
as an exceptional dye in fluorescence detection. QDs-based biosensors
have been described in the literature; however, few of them have truly
progressed to widespread clinical usage. In this work, a chemiluminescent
homogeneous detecting biosensor is fabricated using QDs-doped polystyrene
nanospheres to sensitively detect biomarkers in low-volume serum samples.
Phthalocyanine-dyed and QDs-encapsulated carboxylate-functionalized
polystyrene nanospheres with surface carboxyl groups (PPs and QPs,
respectively) were fabricated and served as triggers and fluorescent
probes, respectively, in this biosensing system. In this sandwich-format
immunoassay, the PPs produced singlet oxygen once sensitized by 680
nm diode lasers, and the QPs, conjugated with antibodies, and then
reacted with the singlet oxygen in the presence of specific antigens
and emitted anti-Stokes fluorescence with wavelengths around 605 nm,
as a result of fluorescence resonance energy transfer (FRET) within
the QPs. We demonstrated the determination of carcinoembryonic antigen
as a model protein target in 25 μL of serum samples with an
unprecedented detection limit of 2.56 × 10–13 M (46 pg/mL) using this biosensor. Furthermore, excellent correlations
(R
2 = 0.99718, n = 107)
were obtained between utilizing this biosensor and commercialized
chemiluminescence immunoassay kits in clinical serum detection. These
results demonstrate that our flexible and reliable biosensor is suitable
for direct integration into clinical diagnostics, and it is expected
to be a promising diagnostic tool for early detection and screening
tests as well as prognosis evaluation for patients.