pubs.acs.

org/JACS Article

Specific Screening of Prostate Cancer Individuals Using an Enzyme-

Assisted Substrate Sensing Platform Based on Hierarchical MOFs
with Tunable Mesopore Size
Liwei Zhao, Jian Yang, Ming Gong, Ke Li, and Jinlou Gu*
Cite This: J. Am. Chem. Soc. 2021, 143, 15145−15151 Read Online

ACCESS Metrics & More Article Recommendations *

sı Supporting Information
See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.
Downloaded via NORTHWEST A & F UNIV on September 27, 2021 at 04:18:09 (UTC).

ABSTRACT: Rapid and specific identification of tumor metabolic

markers is of great significance. Herein, a convenient, reliable and
specific strategy was proposed to screen prostate cancer (PCa)
individuals through indirectly quantifying sarcosine, an early
indicator of PCa, in the clinical urine samples. The success roots
in the rational design of a cascade response model, which takes
integrated sarcosine oxidase (SOX) as a specific recognition unit and
oxygen-sensitive molecule as a signal reporter. The newly developed
hierarchical mesoporous Zr-based metal−organic frameworks with
continuously tunable mesopore size ensure the synergetic work of
the SOX and response unit spatially separated in their neighboring
mesoporous and microporous domains, respectively. The large
mesopore up to 12.1 nm not only greatly enhances the loading
capacity of SOX but also spares enough space for the free diffusion of sarcosine. On this basis, the probe is competent to specifically
check out the tiny concentration change of sarcosine in the urine sample between PCa patients and healthy humans. Such a concept
of enzyme-assisted substrate sensing could be simply extended by altering the type of immobilized enzymes, hopefully setting a
guideline for the rational design of multiple probes to quantify specific biomarkers in complex biological samples.

■ INTRODUCTION
Prostate cancer (PCa) is one of the most common cancers that
The key to our success roots in the design of a cascade
response model, which takes integrated enzyme as a specific
seriously influences the health of men. For most patients, there recognition unit and coupled luminescent molecule as a signal
reporter (Scheme 1). The specificity of an enzymatic reaction
are no obvious clinical symptoms in incipient stages, so that
between sarcosine oxidase (SOX) and sarcosine could
early screening of PCa is crucial for improving their life quality
effectively avoid interference from structurally similar bio-
and reducing mortality.1 Herein, a convenient, reliable, and
molecules. Meanwhile, sarcosine would be effectively con-
specific strategy has been successfully proposed to screen PCa
verted to the products through consuming equimolar dissolved
individuals through quantifying sarcosine levels in the clinical
oxygen (DO). The variation of DO could be immediately
urine samples. Compared to the traditional prostate-specific
distinguished by the coupled oxygen-sensitive indicators,
antigen test to identify PCa, no complex and time-consuming
thereby giving an indirect luminescent response toward
sampling process and no expensive automated equipment are
sarcosine.8,9 Unfortunately, given the poor stability of enzymes
needed hereof.2,3 Sarcosine was verified to be a clinical marker
and the self-aggregation tendency of hydrophobic luminescent
for the early stage PCa, and its levels in urine allow for simple
molecules, the heterogeneous support is essential to couple
and noninvasive early diagnosis.4,5 Generally, sarcosine is them into one entity, thus achieving convenient measurement
produced at the concentration of 1−3 × 10−6 mol·L−1 in of sarcosine in complex biological samples.
human urine under normal physiological conditions, while it Thanks to the open mesochannels and designable topology,
only increases by less than an order of magnitude during the full advantages of hierarchical mesoporous Zr-based metal−
pathological process of PCa.5,6 Additionally, the probes are
generally vulnerable to numerous interferences from possibly
coexistent biomolecules in the clinical samples.7 It is Received: June 2, 2021
consequently hard to precisely distinguish the tiny concen- Published: September 8, 2021
tration change of sarcosine, leading to the risk of false-positive/
negative diagnosis. These issues make sensitive and specific
detection of sarcosine in clinical samples challenging but of
great significance.

© 2021 American Chemical Society https://doi.org/10.1021/jacs.1c05674

15145 J. Am. Chem. Soc. 2021, 143, 15145−15151
Journal of the American Chemical Society pubs.acs.org/JACS Article

Scheme 1. Schematic Illustration for the Proposed Enzyme-
Promoted Cascade Sensory Modela
a mesopore size was applied to specifically screen PCa patients
The enzyme immobilized in the mesoporous domain would rapidly
recognize the biomarker (as its substrate) and convert it into the
corresponding products through an oxygen consumption process
(blue region). Thus, the decreased concentration of O2 would induce
the phosphorescent enhancement of the O2-sensitive TCPP-Pt
strutted in the neighboring microporous domain (pink region),
thereby achieving the specific recognition of the biomarker.
type MOFs (HMUiO) with tailorable mesopore size. The
PEO-PPO-PEO triblock copolymer was utilized as a template
to guide the growth of mesoMOFs with the aid of Hofmeister
salting-in ion, which not only enhances the solubility of organic
linkers but also bridges the precursor of MOFs and PEO-
containing surfactant.31 To regulate the mesopore size of
MOFs, a hydrophobic compound was introduced and
positioned in the micelle core of the surfactant in virtue of
their similar hydrophobicity.32,33 As expected, the mesopore
size could be systematically tuned in the range from 6.4 to 12.1
nm. The correlations of the enzyme loading capacity and
sensory properties of the obtained HMUiO with different
mesopore size were also exploited. The probe with optimized
through quantifying the tiny concentration change of sarcosine
in the urine samples. By altering the type of encapsulated
enzymes, such a concept of enzyme-assisted substrate sensing
could be facilely extended, hopefully paving the way for the
rational design of a variety of other luminescent probes for the
convenient quantification of various biomarkers.

organic frameworks (mesoMOFs) could be taken for

constructing such a heterogeneous system.10−20 Their rigid
and stable mesochannels could work as exoskeletons to
immobilize and protect natural SOX.21−28 Meanwhile, the
phosphorescent carboxylated metalloporphyrin of TCPP-Pt
could serve as colinker to be well incorporated into the
surrounding microporous walls as oxygen-sensitive signal units
(Scheme 1).29 Metallic nodes in MOFs could effectively isolate
TCPP-Pt, thus avoiding the aggregation-induced quenching
effect.8 Additionally, the enzymatic mesopores are seamlessly
surrounded by TCPP-Pt decorated micropores, affording
effective coupling and mass communication between differ-
ent-sized pore units.30 However, the continuous and accurate
expansion of the large mesopore has not been realized in
mesoMOFs up to now, which is a prerequisite to match with
the dimensions of SOX and simultaneously to spare enough
space for the free diffusion of sarcosine to approach enzymes.
Keeping this in mind, we present a simple and reproducible
approach for the synthesis of hierarchical mesoporous UiO-66
■ RESULTS AND DISCUSSION
In a typical synthesis, F127 (PEO106PPO70PEO106) was
selected to direct the growth of mesoMOFs through the
mediating effect of Hofmeister ion of ClO4−.31 To enlarge the
radius of F127 micelle and consequently to regulate the
mesopore size of mesoMOFs, different amounts of hydro-
phobic 1,3,5-trimethylbenzene (TMB) were fed into the initial
reactant as a swelling agent (Figure 1A). The sectional radius
of the complex micelles correlates positively with the ratio of
TMB/F127, agreeing with the fact that TMB could effectively
expand the radius of F127 micelle (Figure S1). The TMB/
F127 complex micelle further works as a directing agent to
produce mesopores with continuously tailorable size. Mean-
while, the O2-sensitive luminescent unit of TCPP-Pt was
compartmentalized into the microporous walls of MOFs
through the one-pot assembly of TCPP-Pt@HMUiO using
BDC-NH2 and TCPP-Pt as colinkers (Figure 1A).29 The
nanoscale TCPP-Pt@HMUiO were formed at various feed

Figure 1. (A) Schematic illustration for the one-pot assembly process of the hierarchical TCPP-Pt@HMUiO with tunable mesopore size through a
Hofmeister ion-mediated template strategy. (B) Nitrogen sorption isotherms and (C) their corresponding BJH pore size distribution profiles of the
TCPP-Pt@HMUiO synthesized with varied TMB/F127 feed ratios of (a) 0, (b) 4.5, (c) 9.1, (d) 18.1, (e) 27.2, and (f) 36.2. (D) The effect of
TMB content in the initial reactant mixture on the distribution of mesopore size in the final product.

15146 https://doi.org/10.1021/jacs.1c05674
J. Am. Chem. Soc. 2021, 143, 15145−15151
Journal of the American Chemical Society
ratios of TMB/F127 as verified by dynamic light scattering
(DLS) measurements (Figure S2).
The topological structures of all samples were well preserved
compared to that of pristine UiO-66 as verified by X-ray
diffraction (XRD), indicating that the introduction of TMB
and TCPP-Pt would not deteriorate crystal walls (Figure S3).34
The N2 sorption isotherms, Brunauer−Emmett−Teller surface
area fitting curves, and pore size distributions of each sample
were measured in detail (Figures S4−S9). The combination of
typical type I and type IV isotherms was obtained from the
nitrogen sorption measurements (Figure 1B). The rising
sorptions at low relative pressure (P/P0 = 0−0.05) could be
attributed to the intrinsic abundant micropores of UiO-66,
while the verticals at high pressure (P/P0 = 0.9−1) were
assigned to the interparticle spaces (embodied as a 70−80 nm
pore size distribution among the nanoparticles (NPs)) (Figure
1C). Notably, along with the increase of TMB/F127, the
position of the capillary condensation steps gradually shifted to
the higher relative pressure (P/P0 varies from 0.82 to 0.88),
corresponding to the continuous enlargement of the mesopore.
The pore diameter was determined by the Barrett−Joyner−
Halenda (BJH) model using the adsorption branch (Figure
1C). A relatively larger mesopore size of 12.1 nm in TCPP-
Pt@HMUiO could be acquired at a high TMB/F127 ratio of
36.2, which is almost twice larger than that (6.4 nm) of TCPP-
Pt@HMUiO synthesized without the addition of TMB.
Further increase the dosing amount of TMB would not
obviously increase the mesopore size. The nonlinear relation-
ship between TMB/F127 feed ratio and TCPP-Pt@HMUiO
mesopore size was fitted (Figure 1D), which is of great
significance for the construction of mesoMOFs with different
mesopore size to satisfy different applications. Combined with
their high surface areas and large pore volumes as well as
tunable pore sizes (Table S1), the as-synthesized TCPP-Pt@
HMUiO could serve as ideal candidates for the immobilization
of diverse enzymes and offers enough space and accessibility
for the communication between enzymes and their substrates.
The gradual expansion of the mesopores can be clearly
observed from the scanning electron microscopy (SEM)
images (Figures 2A−C and S10). Meanwhile, the spherical
NPs with a mean particle size of ca. 100 nm are visible, and
their morphologies are independent of the amount of TMB
introduced. The particle size determined by SEM is slightly
smaller than that calculated by DLS, which is reasonable since
a hydrodynamic layer is usually included in the DLS
measurement. Transmission electron microscope (TEM,
Figures 2D−F and S11) and scanning TEM (STEM) images
(Figure 2G, leftmost panel) further certify the presence of
highly open mesopores in the whole NPs and the pore size
exhibits positive relationship with the introduced amount of
TMB in agreement with the N2 sorption deduction and SEM
images.
The in situ introduction of TCPP-Pt in the microporous wall
of MOFs makes it possible to serve as a cascade signal reporter.
The elemental mappings of the synthesized TCPP-Pt@
HMUiO demonstrate the homogeneous distribution of Zr,
Pt, C, N, and O elements, affirming the expected implantation
of TCPP-Pt into the frameworks (Figure 2G). Furthermore,
the UV−vis absorption spectrum of TCPP-Pt@HMUiO
displays a combination absorbance of aminobenzene at 330
nm and TCPP-Pt at 401 (Soret band), 512 (Q-band), and 538
nm (Q-band) (Figure S12).35,36 The real content of TCPP-Pt
in HMUiO was determined to be around 0.055 mmol per
15147
pubs.acs.org/JACS Article
Figure 2. (A−C) SEM and (D−F) TEM images of the TCPP-Pt@
HMUiO mesoporous nanospheres prepared with different TMB/
F127 feed ratios of (A, D) 0, (B, E) 9.1, and (C, F) 36.2. (G) STEM
image (leftmost panel) and element mappings of Zr, Pt, C, N, and O
for the as-synthesized TCPP-Pt@HMUiO.
gram through a standard curve method (Figure S13). The Pt
content of the TCPP-Pt@HMUiO was quantified to be 0.053
mmol/g by inductively coupled plasma, which is consistent
with the result deduced from the UV−vis measurement.
Meanwhile, the molar ratio of TCPP-Pt to BDC-NH2 was
quantified to be approximately 0.014 in TCPP-Pt@HMUiO
through the liquid 1H NMR spectroscopy analysis (Figures
S14−S16). Encouragingly, the strut of TCPP-Pt on the
micropore wall could effectively avoid its aggregation
quenching in aqueous phase.8 Meanwhile, the separated
luminescent units on micropore wall could form a cascade
response system with enzymes accommodated in the adjacent
mesopores, avoiding the undesirable interaction and config-
uration change between enzymes and luminescent units.
In such heterogeneous sensing platform, the high loading of
SOX is crucial for the rapid conversion and sensitive
recognition of substrates. To achieve this goal, MOFs with
different mesopore size were incubated with natural SOX. As
the mesopore size increased from 6.4 to 12.1 nm, the loading
capacities of SOX significantly increased from 41 to 187 mg
g−1, determined through the standard curve (Figures 3A and
S17). SEM and TEM images were taken to observe the
structural evolvement of TCPP-Pt@HMUiO after enzyme
loading. As shown in Figure S18, the open mesopores are
perfectly preserved on the surface of TCPP-Pt@HMUiO NPs,
indicating that the enzyme would not block the mesoporous
windows on MOFs. TEM images further confirm that the
connected mesoporous channels remain intact after loading
SOX (Figure S19). Meanwhile, the sharp reductions in the
porosity of MOFs further demonstrate that the enzyme is
successfully entrapped within the mesopores rather than
adsorbed on the external surface (Figures S20−S25 and
Table S2). Furthermore, the largest pore volume reduction was
observed for the mesoMOFs with pore size of 12.1 nm.
Obviously, the larger mesopore is beneficial to the high loading
of enzyme. Since the three-dimensional parameters of SOX are
6.9 nm × 7.3 nm × 7.4 nm,6 enough space would reserve for
https://doi.org/10.1021/jacs.1c05674
J. Am. Chem. Soc. 2021, 143, 15145−15151
Journal of the American Chemical Society
Figure 3. (A) The effect of mesopore sizes of the TCPP-Pt@HMUiO
on the loading capacity of SOX therein. (B) The evolvement of the
luminescent spectra of the cascade sensing platform at different
sarcosine concentrations. (C) Plot of luminescent intensity against
sarcosine concentration and the corresponding linear regression
curve. (D) The luminescent response of TCPP-Pt/SOX@HMUiO
with different mesopore size toward sarcosine under oxygenous and
hypoxic conditions within the same testing time.
the free diffusion of sarcosine toward SOX immobilized in
HMUiO with pore size of 12.1 nm.
In the presence of TCPP-Pt/SOX@HMUiO, the sarcosine
would be specifically recognized by the immobilized SOX and
converted into glycine, formaldehyde, and H2O2 through
consuming equamolar DO.37 Since the DO is the main driving
force of the SOX-induced reaction occurring in the
mesoporous region, the decreased oxygen level would give
rise to the optical signal change of TCPP-Pt in the surrounding
microporous walls. As illustrated from the response kinetics
(Figure S26), the luminescent intensity of the sensor gradually
increased upon prolonging the cultivation time and reached
saturation at approximately 60 min, which was set as an
equilibrium time in the subsequent assays for the sarcosine
detecting. Titration experiments displayed that the luminescent
intensity of TCPP-Pt/SOX@HMUiO probe progressively
increased with the addition of sarcosine from 0 to 110 μM
(Figure 3B). The enhanced luminescent intensity presented a
good linear relationship with the sarcosine level in the range of
0−100 μM (R2 = 0.995, Figure 3C). The limit of detection
(LOD) of probe is calculated to be 2.1 μM using the 3σ
IUPAC criteria (3σ/slope), matching well with the concen-
tration window of sarcosine in the urine of PCa patients.6 As a
control experiment, free TCPP-Pt and SOX in the absence of
HMUiO were adopted for the sensing of sarcosine. As shown
in Figure S27, characteristic luminescent peak of the free
TCPP-Pt in the range of 600 to 750 nm is invisible, suggesting
that it was difficult to measure the luminescent properties of
the hydrophobic TCPP-Pt due to its aggregation tendency in
water. After the addition of 100 μM sarcosine into the TCPP-
Pt/SOX system, no luminescent enhancement could be
detected. Therefore, HMUiO played a decisive role as porous
support for the immobilization of enzymes and isolation
TCPP-Pt molecules, thus avoiding the aggregation-induced
quenching effect of hydrophobic TCPP-Pt.
15148
pubs.acs.org/JACS Article
To verify the cascade response mechanism, the sensing
feature of probes with various mesopore sizes was further
investigated under oxygenous and hypoxic conditions (Figure
3D). In the presence of DO, the luminescence response
amplitude of probe significantly increased for the detection of
sarcosine (100 μM) upon expanding mesopore sizes of MOFs
from 8.3 to 12.1 nm. The larger mesopore not only facilitates
the diffusion of sarcosine but also favors more SOX to be
immobilized (Figure 3A), thus more DO would be consumed,
rationalizing the strongest luminescent intensity from probe
with pore size of 12.1 nm. On the other hand, almost no
luminescence change was found when the probes were applied
in the absence of oxygen, no matter how much SOX has been
immobilized (Figure 3D, right panel). Therefore, the DO in
the detected environment plays a decisive role in the cascade
recognition of sarcosine.
To gain an intuitive insight into the interaction between DO
consumed by enzymatic reaction and oxygen-sensitive TCPP-
Pt, an oxygen/nitrogen mixture was introduced into the
sensing system for the regulation of DO levels to simulate the
oxygen-consumption process catalyzed by SOX. The probe
exhibited a weak phosphorescent emission at 665 nm under
the rich oxygen atmosphere. Meanwhile, the peak intensity
significantly increased upon reducing DO concentration
(Figure S28), which resembled the luminescence enhancing
trend resulted from the oxygen consumption during sarcosine
sensing assisted by SOX. These results coincide well with our
proposal that the sarcosine substrate of SOX could be
indirectly quantified through detecting the consumption of
DO (Scheme 1).
To reflect its good reproducibility, the luminescence
variation was measured by alternately adding sarcosine or air
into the suspension of TCPP-Pt/SOX@HMUiO. After an air-
blowing procedure, the luminescence from the probe
recovered to its initial intensity (Figure S29). After four cycles
of experiments, higher than 98% of its first cycling luminescent
intensity could still be maintained. Upon exposure to several
possibly coexistent interfering molecules (including glycine,
tyrosine, histidine, alanine, serine, and uric acid),38 its
luminescent intensity remained almost unaffected (Figure
4A,B). After soaking the as-synthesized TCPP-Pt/SOX@
HMUiO in HEPES buffer solutions for 24 h, no leakages of
SOX and phosphorescent TCPP-Pt were detected (Figures
S30 and S31). Meanwhile, the enzyme and TCPP-Pt molecules
are firmly immobilized in the porous support which could
effectively avoid their leakage in the sensing process (Figures
S32 and S33). The topological and sensory features proceed
with no change even after storing the materials in a refrigerator
for as long as 15 days (Figures S34 and S35). Taking its
excellent specificity together with the high stability and long-
term sensitivity, TCPP-Pt/SOX@HMUiO is competent for
the specific detection of sarcosine in real human urine.
The practicability of the probe was validated by the specific
detection of biomarker in real urine samples (Ruijing Hospital,
Shanghai) from PCa individuals and healthy humans. The new
calibration curve of probe was carried out in urine matrix
before each measurement (Figures S36 and S37). Then, the
levels of sarcosine in urine samples of six healthy volunteers
(including 3 males and 3 females) and three PCa patients were
quantified by using the TCPP-Pt/SOX@HMUiO probe. It
could be observed that the urine samples from PCa patients
result in a significant increase in the luminescent intensity of
the probe compared with those of the healthy human groups
https://doi.org/10.1021/jacs.1c05674
J. Am. Chem. Soc. 2021, 143, 15145−15151
Journal of the American Chemical Society
Figure 4. Changes of the luminescent intensity of the TCPP-Pt/
SOX@HMUiO at 665 nm against various interfering species (0−100
μM) (A) without and (B) with the addition of sarcosine. Sar,
sarcosine; Gly, glycine; Tyr, tyrosine; His, histidine; Ala, alanine; Ser,
serine; and UA, uric acid. (C) The emission intensity of TCPP-Pt/
SOX@HMUiO for three PCa patients (red columns), three healthy
males (green columns), and three healthy females (blue columns).
(D) Luminescent spectra of TCPP-Pt/SOX@HMUiO from human
urine added with different concentrations of sarcosine.
(Figure 4C), indicating that the probe could effectively
respond to sarcosine in the urine samples. By converting the
luminescent intensity of probe into the biomarker concen-
tration, the mean sarcosine concentration of patient samples
was determined to be around 16.9 μM, which was
approximately 8 times higher than that of a healthy human.
These results indicated that the developed probe could
effectively and precisely provide a positive diagnosis of
sarcosine biomarker for PCa patients. To further evaluate the
accuracy of the procedure, the recovery experiments were
carried out on the urine sample of patient 1 through the extra
addition of sarcosine.39 As shown in Figure 4D, the
luminescent intensity of TCPP-Pt/SOX@HMUiO probe
increased upon adding the urine sample of patient 1 to the
sensory system, and the concentration of sarcosine was
quantified to be about 21.25 μM. When extra sarcosine (30
μM) was added, a stronger luminescent enhancement could be
observed (Figure 4D, blue line), and the calculated sarcosine
concentration matched well with the extra added amount of
sarcosine. The high recoveries ranging from 98.91% to
101.94% evidence the accuracy of the probe for the detection
of sarcosine in the urine sample (Table S3), further prefiguring
its great practical prospect in screening of PCa patients from a
large group of asymptomatic healthy or diseased population.
Based on such a cascade sensory model, other metabolic
biomarkers could be conveniently checked as long as the
immobilized enzymes were altered. Taking the representative
enzyme of glucose oxidase (GOX) as an example, the loading
capacity of GOX in the TCPP-Pt@HMUiO probe with
mesopore size of 12.1 nm could reach high up to
approximately 214 mg·g−1 (Figure S38). The elaborated
TCPP-Pt/GOX@HMUiO presented a good linear response
toward the detection of glucose (Figures S39 and S40), and
the LOD value was determined to be about 2.7 μM. Further
extension of such sensory system could focus on the
immobilization of larger-size enzymes such as urate oxidase
15149
pubs.acs.org/JACS Article
(UOX), and some chiral-related enzymes such as L/D-amino
acid oxidase (AOX). Some typical enzyme/substrate pairs (i.e.,
GOX/glucose, AOX/amino acid, and ChOX/cholesterol as
well as UOX/uric acid) are listed in Table S4.
■ CONCLUSIONS
In summary, an enzyme-assisted substrate sensing model was
proposed for the specific screening of PCa patients. Newly
developed hierarchical mesoMOFs with a continuously tunable
mesopore size between 6.4 and 12.1 nm guaranteed the
synergetic work of the encapsulated SOX and integrated
oxygen-sensitive indicator. The proposed probe could realize a
linear luminescent quantification of sarcosine in the range of
0−100 μM with a LOD of 2.1 μM. Based on the sensitivity and
specificity of the enzymatic reaction, the indirect identification
of tiny sarcosine changes in the urine samples between PCa
individuals and healthy humans was successfully realized. Such
hierarchical mesoMOFs are competent for the immobilization
of various oxidases as recognition units, thus holding great
promise to quantify a diversity of biomarkers for the diagnosis
of relative diseases.
■
ASSOCIATED CONTENT
* Supporting Information
sı
The Supporting Information is available free of charge at
https://pubs.acs.org/doi/10.1021/jacs.1c05674.
Experimental details, DLS profiles, XRD patterns, SEM
and TEM images of the mesoMOFs with different TMB,
1
H NMR, luminescent spectra, UV−vis spectra, and
Tables S1−S4 of the mesoMOFs (PDF)
■
AUTHOR INFORMATION
Corresponding Author
Jinlou Gu − Key Laboratory for Ultrafine Materials of
Ministry of Education, School of Materials Science and
Engineering, East China University of Science and
Technology, Shanghai 200237, China; orcid.org/0000-
0002-3190-573X; Email: jinlougu@ecust.edu.cn
Authors
Liwei Zhao − Key Laboratory for Ultrafine Materials of
Ministry of Education, School of Materials Science and
Engineering, East China University of Science and
Technology, Shanghai 200237, China
Jian Yang − Key Laboratory for Ultrafine Materials of
Ministry of Education, School of Materials Science and
Engineering, East China University of Science and
Technology, Shanghai 200237, China; orcid.org/0000-
0002-4451-0497
Ming Gong − Key Laboratory for Ultrafine Materials of
Ministry of Education, School of Materials Science and
Engineering, East China University of Science and
Technology, Shanghai 200237, China
Ke Li − Key Laboratory for Ultrafine Materials of Ministry of
Education, School of Materials Science and Engineering, East
China University of Science and Technology, Shanghai
200237, China
Complete contact information is available at:
https://pubs.acs.org/10.1021/jacs.1c05674
Notes
The authors declare no competing financial interest.
https://doi.org/10.1021/jacs.1c05674
J. Am. Chem. Soc. 2021, 143, 15145−15151
Journal of the American Chemical Society
■
ACKNOWLEDGMENTS
This work was financially supported by the Natural Science
Foundation of China (21975072, 51902106) and the Natural
Science Foundation of Shanghai (18ZR1408700).
■ REFERENCES
(1) Berger, M. F.; Lawrence, M. S.; Demichelis, F.; Drier, Y.;
Cibulskis, K.; Sivachenko, A. Y.; Sboner, A.; Esgueva, R.; Pflueger, D.;
Sougnez, C.; Onofrio, R.; Carter, S. L.; Park, K.; Habegger, L.;
Ambrogio, L.; Fennell, T.; Parkin, M.; Saksena, G.; Voet, D.; Ramos,
A. H.; Pugh, T. J.; Wilkinson, J.; Fisher, S.; Winckler, W.; Mahan, S.;
Ardlie, K.; Baldwin, J.; Simons, J. W.; Kitabayashi, N.; MacDonald, T.
Y.; Kantoff, P. W.; Chin, L.; Gabriel, S. B.; Gerstein, M. B.; Golub, T.
R.; Meyerson, M.; Tewari, A.; Lander, E. S.; Getz, G.; Rubin, M. A.;
Garraway, L. A. The Genomic Complexity of Primary Human
Prostate Cancer. Nature 2011, 470, 214−220.
(2) Lilja, H.; Ulmert, D.; Vickers, A. J. Prostate-Specific Antigen and
Prostate Cancer: Prediction, Detection and Monitoring. Nat. Rev.
Cancer 2008, 8, 268−278.
(3) Li, H.; Huang, Y.; Hou, G.; Xiao, A.; Chen, P.; Liang, He.;
Huang, Y.; Zhao, X.; Liang, L.; Feng, X.; Guan, B.-O. Single-Molecule
Detection of Biomarker and Localized Cellular Photothermal Therapy
Using an Optical Microfiber with Nanointerface. Sci. Adv. 2019, 5,
eaax4659.
(4) Biavardi, E.; Tudisco, C.; Maffei, F.; Motta, A.; Massera, C.;
Condorelli, G. G.; Dalcanale, E. Exclusive Recognition of Sarcosine in
Water and Urine by a Cavitand-Functionalized Silicon Surface. Proc.
Natl. Acad. Sci. U. S. A. 2012, 109, 2263−2268.
(5) Sreekumar, A.; Poisson, L. M.; Rajendiran, T. M.; Khan, A. P.;
Cao, Q.; Yu, J.; Laxman, B.; Mehra, R.; Lonigro, R. J.; Li, Y.; Nyati, M.
K.; Ahsan, A.; Kalyana-Sundaram, S.; Han, B.; Cao, X.; Byun, J.;
Omenn, G. S.; Ghosh, D.; Pennathur, S.; Alexander, D. C.; Berger, A.;
Shuster, J. R.; Wei, J. T.; Varambally, S.; Beecher, C.; Chinnaiyan, A.
M. Metabolomic Profiles Delineate Potential Role for Sarcosine in
Prostate Cancer Progression. Nature 2009, 457, 910−914.
(6) Henderson, C. J.; Pumford, E.; Seevaratnam, D. J.; Daly, R.; Hall,
E. A. H. Gene to Diagnostic: Self Immobilizing Protein for Silica
Microparticle Biosensor, Modelled With Sarcosine Oxidase. Bio-
materials 2019, 193, 58−70.
(7) He, J.; Li, C.; Ding, L.; Huang, Y.; Yin, X.; Zhang, J.; Zhang, J.;
Yao, C.; Liang, M.; Pirraco, R. P.; Chen, J.; Lu, Q.; Baldridge, R.;
Zhang, Y.; Wu, M.; Reis, R. L.; Wang, Y. Tumor Targeting Strategies
of Smart Fluorescent Nanoparticles and Their Applications in Cancer
Diagnosis and Treatment. Adv. Mater. 2019, 31, 1902409.
(8) Yang, J.; Wang, Z.; Li, Y.; Zhuang, Q.; Gu, J. Real-Time
Monitoring of Dissolved Oxygen with Inherent Oxygen-Sensitive
Centers in Metal-Organic Frameworks. Chem. Mater. 2016, 28,
2652−2658.
(9) Xu, R.; Wang, Y.; Duan, X.; Lu, K.; Micheroni, D.; Hu, A.; Lin,
W. Nanoscale Metal-Organic Frameworks for Ratiometric Oxygen
Sensing in Live Cells. J. Am. Chem. Soc. 2016, 138, 2158−2161.
(10) Wang, S.; Chen, L.; Wahiduzzaman, M.; Tissot, A.; Zhou, L.;
Ibarra, I. A.; Gutiérrez-Alejandre, A.; Lee, J. S.; Chang, J.-S.; Liu, Z.;
Marrot, J.; Shepard, W.; Maurin, G.; Xu, Q.; Serre, C. A Mesoporous
Zirconium-Isophthalate Multifunctional Platform. Matter 2021, 4,
182−194.
(11) Wang, K. Y.; Feng, L.; Yan, T. H.; Qin, J. S.; Li, C. X.; Zhou, H.
C. Morphology Transcription in Hierarchical MOF-on-MOF
Architectures. ACS Mater. Lett. 2021, 3, 738−743.
(12) Li, J.-X.; Chang, G.-G.; Tian, G.; Pu, C.; Huang, K.-X.; Ke, S.-
C.; Janiak, C.; Yang, X.-Y. Near-Linear Controllable Synthesis of
Mesoporosity in Hierarchical UiO-66 by Template-Free Nucleation-
Competition. Adv. Funct. Mater. 2021, 31, 2102868.
(13) Cirujano, F. G.; Martin, N.; Wee, L. H. Design of Hierarchical
Architectures in Metal-Organic Frameworks for Catalysis and
Adsorption. Chem. Mater. 2020, 32, 10268−10295.
(14) Zhang, F.; Hu, X.; Roth, E. W.; Kim, Y.; Nguyen, S. T.
Template-Assisted, Seed-Mediated Synthesis of Hierarchically Meso-
15150
pubs.acs.org/JACS Article
porous Core-Shell UiO-66: Enhancing Adsorption Capacity and
Catalytic Activity through Iterative Growth. Chem. Mater. 2020, 32,
4292−4302.
(15) Yang, Y.; Arqué, X.; Patiño, T.; Guillerm, V.; Blersch, P.-R.;
Pérez-Carvajal, J.; Imaz, I.; Maspoch, D.; Sánchez, S. Enzyme-
Powered Porous Micromotors Built from a Hierarchical Micro- and
Mesoporous UiO-Type Metal-Organic Framework. J. Am. Chem. Soc.
2020, 142, 20962−20967.
(16) Chen, Y.; Li, P.; Zhou, J.; Buru, C. T.; Đorđević, L.; Li, P.;
Zhang, X.; Cetin, M. M.; Stoddart, J. F.; Stupp, S. I.; Wasielewski, M.
R.; Farha, O. K. Integration of Enzymes and Photosensitizers in a
Hierarchical Mesoporous Metal-Organic Framework for Light-Driven
CO2 Reduction. J. Am. Chem. Soc. 2020, 142, 1768−1773.
(17) Wang, K.; Feng, L.; Yan, T.; Wu, S.; Joseph, E. A.; Zhou, H.
Rapid Generation of Hierarchically Porous Metal-Organic Frame-
works through Laser Photolysis. Angew. Chem., Int. Ed. 2020, 59,
11349−11354.
(18) Lu, J.; Wu, J.-K.; Jiang, Y.; Tan, P.; Zhang, L.; Lei, Y.; Liu, X.-
Q.; Sun, L.-B. Fabrication of Microporous Metal-Organic Frameworks
in Uninterrupted Mesoporous Tunnels: Hierarchical Structure for
Efficient Trypsin Immobilization and Stabilization. Angew. Chem., Int.
Ed. 2020, 59, 6428−6434.
(19) Li, P.; Chen, Q.; Wang, T. C.; Vermeulen, N. A.; Mehdi, B. L.;
Dohnalkova, A.; Browning, N. D.; Shen, D.; Anderson, R.; Gómez-
Gualdrón, D. A.; Cetin, F. M.; Jagiello, J.; Asiri, A. M.; Stoddart, J. F.;
Farha, O. K. Hierarchically Engineered Mesoporous Metal-Organic
Frameworks toward Cell-Free Immobilized Enzyme Systems. Chem.
2018, 4, 1022−1034.
(20) Chen, Y.; Li, P.; Modica, J. A.; Drout, R. J.; Farha, O. K. Acid-
Resistant Mesoporous Metal-Organic Framework toward Oral Insulin
Delivery: Protein Encapsulation, Protection, and Release. J. Am.
Chem. Soc. 2018, 140, 5678−5681.
(21) Liang, J.; Gao, S.; Liu, J.; Zulkifli, M. Y. B.; Xu, J.; Scott, J.;
Chen, V.; Shi, J.; Rawal, A.; Liang, K. Hierarchically Porous
Biocatalytic MOF Microreactor as a Versatile Platform towards
Enhanced Multienzyme and Cofactor-Dependent Biocatalysis. Angew.
Chem., Int. Ed. 2021, 60, 5421−5428.
(22) Wang, X.; Lan, P. C.; Ma, S. Metal-Organic Frameworks for
Enzyme Immobilization: Beyond Host Matrix Materials. ACS Cent.
Sci. 2020, 6, 1497−1506.
(23) Liang, S.; Wu, X.-L.; Xiong, J.; Zong, M.-H.; Lou, W.-Y. Metal-
Organic Frameworks as Novel Matrices for Efficient Enzyme
Immobilization: An Update Review. Coord. Chem. Rev. 2020, 406,
213149.
(24) Huang, S.; Kou, X.; Shen, J.; Chen, G.; Ouyang, G. Armor-
Plating” Enzymes with Metal-Organic Frameworks (MOFs). Angew.
Chem., Int. Ed. 2020, 59, 8786−8798.
(25) Hu, C.; Bai, Y.; Hou, M.; Wang, Y.; Wang, L.; Cao, X.; Chan,
C.-W.; Sun, H.; Li, W.; Ge, J.; Ren, K. Defect-Induced Activity
Enhancement of Enzyme-Encapsulated Metal-Organic Frameworks
Revealed in Microfluidic Gradient Mixing Synthesis. Sci. Adv. 2020, 6,
eaax5785.
(26) Guo, J.; Yang, L.; Gao, Z.; Zhao, C.; Mei, Y.; Song, Y.-Y. Insight
of MOF Environment-Dependent Enzyme Activity via MOFs-in-
Nanochannels Configuration. ACS Catal. 2020, 10, 5949−5958.
(27) Chen, S.-Y.; Lo, W.-S.; Huang, Y.-D.; Si, X.; Liao, F.-S.; Lin, S.-
W.; Williams, B. P.; Sun, T.-Q.; Lin, H.-W.; An, Y.; Sun, T.; Ma, Y.;
Yang, H.-C.; Chou, L.-Y.; Shieh, F.-K.; Tsung, C.-K. Probing
Interactions between Metal-Organic Frameworks and Freestanding
Enzymes in a Hollow Structure. Nano Lett. 2020, 20, 6630−6635.
(28) Gkaniatsou, E.; Sicard, C.; Ricoux, R.; Benahmed, L.;
Bourdreux, F.; Zhang, Q.; Serre, C.; Mahy, J.; Steunou, N. Enzyme
Encapsulation in Mesoporous Metal-Organic Frameworks for
Selective Biodegradation of Harmful Dye Molecules. Angew. Chem.,
Int. Ed. 2018, 57, 16141−16146.
(29) Sun, Y.; Sun, L.; Feng, D.; Zhou, H.-C. An In Situ One-Pot
Synthetic Approach towards Multivariate Zirconium MOFs. Angew.
Chem., Int. Ed. 2016, 55, 6471−6475.
https://doi.org/10.1021/jacs.1c05674
J. Am. Chem. Soc. 2021, 143, 15145−15151
Journal of the American Chemical Society pubs.acs.org/JACS Article

(30) Li, K.; Yang, J.; Gu, J. Spatially Organized Functional

Bioreactors in Nanoscale Mesoporous MOFs for Cascade Scavenging
of Intracellular ROS. Chem. Mater. 2021, 33, 2198−2205.
(31) Li, K.; Yang, J.; Huang, R.; Lin, S.; Gu, J. Ordered Large-Pore
MesoMOFs Based on Synergistic Effects of TriBlock Polymer and
Hofmeister Ion. Angew. Chem., Int. Ed. 2020, 59, 14124−14128.
(32) Huang, H.; Li, J. R.; Wang, K.; Han, T.; Tong, M.; Li, L.; Xie,
Y.; Yang, Q.; Liu, D.; Zhong, C. An In Situ Self-Assembly Template
Strategy for the Preparation of Hierarchical-Pore Metal-Organic
Frameworks. Nat. Commun. 2015, 6, 8847.
(33) Zhao, D.; Feng, J.; Huo, Q.; Melosh, N.; Fredrickson, G. H.;
Chmelka, B. F.; Stucky, G. D. Triblock Copolymer Syntheses of
Mesoporous Silica with Periodic 50 to 300 Angstrom Pores. Science
1998, 279, 548−552.
(34) Cavka, J. H.; Jakobsen, S.; Olsbye, U.; Guillou, N.; Lamberti,
C.; Bordiga, S.; Lillerud, K. P. A New Zirconium Inorganic Building
Brick Forming Metal Organic Frameworks with Exceptional Stability.
J. Am. Chem. Soc. 2008, 130, 13850−13851.
(35) Zhang, X.; Wasson, M. C.; Shayan, M.; Berdichevsky, E. K.;
Ricardo-Noordberg, J.; Singh, Z.; Papazyan, E. K.; Castro, A. J.;
Marino, P.; Ajoyan, Z.; Chen, Z.; Islamoglu, T.; Howarth, A. J.; Liu,
Y.; Majewski, M. B.; Katz, M. J.; Mondloch, J. E.; Farha, O. K. A
Historical Perspective on Porphyrin-Based Metal-Organic Frame-
works and Their Applications. Coord. Chem. Rev. 2021, 429, 213615.
(36) Chen, J.; Zhu, Y.; Kaskel, S. Porphyrin-Based Metal-Organic
Frameworks for Biomedical Applications. Angew. Chem., Int. Ed. 2021,
60, 5010−5035.
(37) Jiang, X.-Y.; Zhang, L.; Liu, Y.-L.; Yu, X.-D.; Liang, Y.-Y.; Qu,
P.; Zhao, W.-W.; Xu, J.-J.; Chen, H.-Y. Hierarchical CuInS2-Based
Heterostructure: Application for Photocathodic Bioanalysis of
Sarcosine. Biosens. Bioelectron. 2018, 107, 230−236.
(38) Mbage, B.; Li, Y.; Si, H.; Zhang, X.; Li, Y.; Wang, X.; Salah, A.;
Zhang, K. Fabrication of Folate Functionalized Polyoxometalate
Nanoparticle to Simultaneously Detect H2O2 and Sarcosine in
Colorimetry. Sens. Actuators, B 2020, 304, 127429.
(39) Yu, X.; Munge, B.; Patel, V.; Jensen, G.; Bhirde, A.; Gong, J. D.;
Kim, S. N.; Gillespie, J.; Gutkind, J. S.; Papadimitrakopoulos, F.;
Rusling, J. F. Carbon Nanotube Amplification Strategies for Highly
Sensitive Immunodetection of Cancer Biomarkers. J. Am. Chem. Soc.
2006, 128, 11199−11205.

15151 https://doi.org/10.1021/jacs.1c05674
J. Am. Chem. Soc. 2021, 143, 15145−15151