Metal–organic
frameworks (MOFs) have many attractive features,
including tunable composition, rigid structure, controllable pore
size, and large specific surface area, and thus are highly applicable
in molecular analysis. Depending on the MOF structure, a high number
of unsaturated metal sites can be exposed to catalyze chemical reactions.
In the present work, we report that using both Co(II) and Fe(III)
to prepare the MIL-88(NH2) MOF, we can produce the bimetallic
MOF that can catalyze the conversion of 3,3′,5,5″-tetramethylbenzidine
(TMB) to a color product through a reaction with H2O2 at a higher reaction rate than the monometallic Fe-MIL-88(NH2). The Michaelis constants (K
m) of the catalytic reaction for TMB and H2O2 are 3–5 times smaller, and the catalytic constants (k
cat) are 5–10 times higher than those
of the horseradish peroxidase (HRP), supporting ultrahigh peroxidase-like
activity. These values are also much more superior to those of the
HRP-mimicking MOFs reported previously. Interestingly, the bimetallic
MOF can be coupled with glucose oxidase (GOx) to trigger the cascade
enzymatic reaction for highly sensitive detection of extracellular
vesicles (EVs), a family of important biomarkers. Through conjugation
to the aptamer that recognizes the marker protein on EV surface, the
MOF can help isolate the EVs from biological matrices, which are subsequently
labeled by GOx via antibody recognition. The cascade enzymatic reaction
between MOF and GOx enables the detection of EVs at a concentration
as low as 7.8 × 104 particles/mL. The assay can be
applied to monitor EV secretion by cultured cells and also can successfully
detect the different EV quantities in the sera samples collected from
cancer patients and healthy controls. Overall, we prove that the bimetallic
Fe/Co-MIL-88(NH2) MOF, with its high peroxidase activity
and high biocompatibility, is a valuable tool deployable in clinical
assays to facilitate disease diagnosis and prognosis.