Owing to its high throughput, simplicity, and rapidity,
enzyme-linked
immunosorbent assay (ELISA) has attracted much attention in the field
of immunoassays. However, the traditional ELISA usually affords a
single signal readout and the labeling ability of the enzyme used
is poor, resulting in low accuracy and a limited detection range.
Herein, a vanadium nanospheres (VNSs)-mediated competitive ratio nanozymes-linked
immunosorbent assay (VNSs-RNLISA) was created for the sensitive detection
of the T-2 toxin (T-2). As the key to the biosensor, the VNSs with
superoxide dismutase-like and peroxidase-like dual-enzyme mimetic
activities were synthesized by a one-step hydrothermal method, which
oxidized 1,1-diphenyl-2-picryl-hydrazyl fading and catalyzed 3,3′,5,5′-tetramethylbenzidine
(TMB) color development. Therefore, T-2 could not only be qualitatively
measured with the naked eye but also be quantitatively evaluated by
monitoring the ratio of absorbance at 450 and 517 nm wavelengths.
Moreover, the characterization of a VNSs-labeled antibody probe showed
strong dual-enzymatic activity, excellent stability, and high affinity
with T-2 [the affinity constant (k
a) was
approximately 1.36 × 108 M–1], which
can significantly improve the detection sensitivity. The limit of
detection of VNSs-RNLISA was 0.021 ng/mL, which was approximately
27-fold more sensitive than the single signal nanozymes-linked immunosorbent
assay (0.561 ng/mL). Besides, the change in the ratio of absorbance
(Δ450/Δ517) decreased linearly in a range of 0.22–13.17
ng/mL, outperforming the detection range of a single-mode nano-enzyme-linked
immunosorbent assay using TMB by a factor of 1.6 times. Furthermore,
the VNSs-RNLISA was successfully used to identify T-2 in maize and
oat samples, with recoveries ranging from 84.216 to 125.371%. Overall,
this tactic offered a promising platform for the quick detection of
T-2 in food and might broaden the application range of the enzyme-linked
immunosorbent assay.