Herein, an electrochemical (EC)–photoelectrochemical
(PEC)
dual-mode biosensor was constructed for cytokeratin 19 fragment 21-1
(CYFRA21-1) assay based on the dual-signaling electrochemical ratiometric
strategy and “on–off–on” PEC method. The
indium tin oxide (ITO) electrode was modified by 3,4,9,10-perylenetetracarboxylic
dianhydride (PTCDA)@C60 and gold nanoparticles (Au NPs),
and the double-stranded DNA composed of thiol/methylene blue (MB)-labeled
single-stranded DNA (ssDNA) (S0-MB) and antibody/ferrocene (Fc)-labeled
ssDNA (Ab1-S1-Fc) was immobilized on the Au NPs/PTCDA@C60/ITO electrode via the Au–S bond between Au NPs and thiol
of S0-MB. With the help of another antibody-labeled ssDNA (Ab2-S2),
the presence of CYFRA21-1 triggered a typical antigen–antibody
sandwich immune reaction (Ab1, CYFRA21-1, and Ab2) and proximity hybridization
between Ab1-S1-Fc and Ab2-S2. This caused the release of Ab1-S1-Fc
from the modified electrode and the change of S0-MB to a hairpin structure,
resulting in a decrease (an increase) of the oxidation peak current
of Fc (MB) and an increase of the photocurrent due to the enhancing
(inhibiting) effect of MB (Fc) on the photoelectric performance of
the Au NPs/PTCDA@C60/ITO electrode. Thus, CYFRA21-1 was
detected by the developed EC–PEC dual-mode sensing platform
sensitively, and the linear response ranges of 0.001–40 ng/mL
with a detection limit of 0.3 pg/mL for the EC technique and 0.0001–4
ng/mL with a detection limit of 0.03 pg/mL for the PEC method were
obtained. Furthermore, by changing the specific antibodies of disease-related
biomarkers, the developed dual-mode biosensing platform could be readily
extended to detect other antigens, implying its great potential applications
in biological analysis and early disease diagnosis.