Advanced Materials

The endothelialization of organ-on-chip platforms and vascular implants is often limited by slow cell attachment and unstable monolayer formation. This work presents a scalable workflow that imprints micro- and nano-gratings into elastin-like recombinamer (ELR)-based hydrogels, enabling rapid endothelial cell capture and accelerating monolayer formation within 14 days. Three gelatin-ELR formulations are engineered, with {superscript 1}H-NMR confirming incorporation of sequences designed to modulate bioactivity (ELR1: inert; ELR2: uPA-responsive; ELR3: RGD-adhesive). ELR incorporation generates fibrillar microstructures and enhances mechanical performance, yielding elastic-dominant networks suitable for high-fidelity pattern transfer and stable culture. Using this library, the combined effects of ELR bioactivity and groove geometry on human iPSC-derived endothelial cells (iPSC-ECs) are systematically evaluated. In a 15-minute attachment assay, patterned ELR composites markedly improve cell retention compared to gelatin, with ELR2 on [~]350 nm and [~]4 {micro}m grooves performing best, consistent with controlled, cell-mediated interfacial remodeling. This early advantage persists, as ELR2 and ELR3 hydrogels support rapid alignment and reach confluence by day 14, whereas gelatin remains sub-confluent. Cytoskeletal analysis confirms F-actin alignment. By combining enhanced early capture with protease-regulated remodeling, ELR2 identifies a favorable design window. These results establish a materials design framework linking programmable ELR chemistry with surface topography to engineer endothelial interfaces, providing a versatile platform for vascular biomaterials and microphysiological systems.

We present a proof-of-concept platform in which amyloids are displayed on the surface of engineered Bacillus subtilis spores for bioengineered materials. Amyloids possess high tensile strength, elasticity, and tunable assembly, but their use is limited by inaccessible native sources and low-yield or toxic heterologous expression. Here, spores were engineered to display the native amyloid TasA and Humboldt squid suckerins 9 and 10 as fusions to the spore coat protein CotY. Amyloid production and fibril formation were confirmed by Western blot and X-34 staining, and quantitative analysis indicated mg/L-level yields. Atomic force microscopy revealed altered stiffness and surface ultrastructure, and incorporation of amyloid-displaying spores into resin-based 3D printing modified tensile strength. These findings highlight spore-based amyloid display as a scalable, modular platform for materials applications, leveraging established industrial spore production.

The human vasculature is a complex, multiscale system comprising hierarchical networks of macroscale to microscopic vessels. Existing in vitro fabrication techniques often fail to bridge these disparate scales, as high-resolution methods like multiphoton ablation are too slow for replicating larger vessels, while 3D printing lacks the resolution for fine microscale features. Here, we report a "twisted wire templating" strategy capable of generating perfusable bifurcating hydrogel networks that seamlessly transition from the macro- to the micro-scale (2.3 mm to 140 {micro}m) through seven orders of bifurcations. By optimizing wire-twisting geometries and polyurethane dip-coating, we overcame instability-driven bead formation to ensure replication fidelity across the networks. Fabrication rigs were reconfigured from existing 2D planar layouts to 3D reconfigurable architectures to better replicate 3D vessel geometries which simultaneously reducing the laboratory footprint and fabrication times by 47%. Using a Taguchi orthogonal array, we further optimized surface chemistry and hydrogel composition to inhibit structural failure during template extraction, resulting in fully patent, perfusable networks. This method provides a robust, low-cost, and scalable foundation for creating physiologically representative vascular models for investigating multiscale disease mechanisms and organ-level tissue engineering.

Direct ink writing is compatible with an expansive materials palette. While enabling diverse applications, this materials versatility brings significant bottlenecks in ink formulation, often requiring the mixing, printing, and testing of dozens to hundreds of ink compositions over the course of a project. To accelerate ink-space exploration, we introduce gradient embedded multinozzle (GEM) printheads that combine the high-throughput parallelized printing of multinozzles with combinatorial ink mixing. These printheads allow simultaneous mixing of two-, three-, and four-input inks which are distributed to printer nozzles to create complex 3D structures with graded compositions of inks. Using a two-way GEM printhead, we vali-date cell compatibility by printing scaffolds containing various concentrations of fibroblasts and observing non-linear compaction behaviours. We next test a three-way GEM multinozzle to print ten compositions of di- and multi-functionalized poly(ethylene-glycol) diacrylate hydrogel tri-leaflet valves, optimizing for stiffness, swelling ratio, and toughness. Our GEM multinozzles are compatible with open-source printers and either pressure- or volume-driven extrusion systems and promise to accelerate iterative ink design and testing.

Ingestible electronics enable the tracking and treatment of gastrointestinal and systemic diseases. However, bulky batteries and circuit boards require large capsules that can result in bowel obstruction, a medical emergency. Here, we engineered a 9 x 26 mm electronic pill capable of triggered severing into tiny pieces with sizes clinically proven to reduce obstruction risk. Our capsule enables multicomponent circuit boards to connect with separately encapsulated powering elements via conductive, interlocking connections. Heat induced softening of polyethylene glycol/polycaprolactone channels activates a spring to separate encapsulated components into inert 9 x 15 mm segments, facilitating intestinal passage. Separation triggers included closed-loop sensors and time-delay circuits. In vivo swine studies demonstrate the ability of our capsules to sense luminal oxygen changes via an optoelectronic sensor, locally trigger upadacitinib delivery, and facilitate safe excretion.

Extracellular matrix (ECM) mechanical properties regulate tissue homeostasis and disease progression, with persistent ECM stiffening serving as a hallmark of fibrosis; yet, the early transition from healthy to diseased tissue remains poorly understood. Dynamic three-dimensional (3D) tissue models that capture early-stage stiffening are needed to investigate cellular responses during disease initiation. This work presents an innovative platform for studying cell responses in 3D environments undergoing active matrix stiffening. A bioinspired synthetic ECM incorporates collagen-mimetic peptides and employs sequential, non-terminal strain-promoted azide-alkyne cycloaddition (SPAAC) reactions to enable controlled increases in matrix stiffness over physiologically relevant timescales. Alternating polymer incubations produce a 2.5-fold increase in storage modulus over 72 hours, modeling the mechanical transition from healthy to early-stage fibrotic lung tissue. Live-cell reporter fibroblasts enable real-time monitoring of alpha-smooth muscle actin (SMA) expression, revealing significant upregulation during matrix stiffening that remains transient and difficult to detect via traditional endpoint assays. Active stiffening also modulates fibroblast motility, transiently increasing migration speed while persistently enhancing directional persistence. Complementary computational reaction-diffusion modeling provides mechanistic insight into modulus gradient formation and reaction kinetics. This versatile toolbox enables investigation of early mechanobiological responses to matrix stiffening and may aid identification of markers of fibrotic disease onset.

Clinical outcomes in aggressive breast cancer vary widely, in part because the tumor microenvironment is structured to exclude immune infiltration. Low antigen load, dysfunctional antigen-presenting cells, T cell exclusion and exhaustion, and a stiff extracellular matrix that physically restricts immune cell trafficking work together to form a suppressive barrier that current immunotherapies struggle to overcome. We addressed this barrier using ultrasound (US)-activated nanobubbles (NBs), a drug-free intervention based on perfluoropropane-filled nanoparticles. The size and deformable phospholipid shell enable NBs to achieve deep tumor penetration and a uniform distribution throughout the entire tumor. Upon ultrasound activation, NBs generate localized mechanical forces that restore extracellular matrix elasticity, disrupt tumor transport barriers, and drive HMGB1 release, re-engaging endogenous antitumor immunity without pharmacological agents. In a syngeneic triple-negative breast cancer model, US-NB treatment depleted immunosuppressive myeloid cells 3-fold within 3 hours, followed by a greater than 5-fold increase in the ratio of antigen-experienced to suppressive T cells at 48 hours. US-NB drives rapid infiltration of CD4+ and CD8+ T cells within 48 hours. US-NB treatment achieved an 85% cure rate in the D2A1 model; cured animals maintained durable systemic immune memory, rejecting both local and systemic tumor rechallenge. Consistent therapeutic benefit was observed in a luminal B-like mammary tumor model (E0771), supporting activity across breast cancer subtypes. These results establish US-NB mechanical immunomodulation as a drug-free therapeutic strategy capable of generating robust and durable antitumor immunity, acting through biophysical tissue properties rather than tumor-specific molecular targets. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=90 SRC="FIGDIR/small/714247v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@b1ed5forg.highwire.dtl.DTLVardef@1572a98org.highwire.dtl.DTLVardef@1ad6906org.highwire.dtl.DTLVardef@1ca1b36_HPS_FORMAT_FIGEXP M_FIG C_FIG

The clinical translation of engineered skeletal muscle (eSM) for volumetric muscle regeneration is hindered by the challenge of establishing a functional vascular network capable of sustaining its high metabolic demand and ensuring graft survival. Here, we present a bottom-up biofabrication strategy to generate a pre-vascularized in vitro eSM model through the modular assembly of independently matured muscle and vascular compartments. C2C12 myoblasts were encapsulated within core-shell fibers using rotary wet-spinning (RoWS), yielding anisotropically aligned, multinucleated, and contractile myofibers expressing myosin heavy chain and sarcomeric -actinin. In parallel, gelatin methacryloyl (GelMA)-based microvascular seeds ({micro}VS), pre-endothelialized with human umbilical vein endothelial cells, were engineered to guide rapid and structurally stable vascular formation while preventing uncontrolled capillary self-organization. Fully endothelialized {micro}VS were incorporated into a pro-angiogenic bioink and processed via RoWS to generate tubular vascular fibers with physiological diameters (100-200 m) and continuous CD31-positive lumens. After independent maturation, muscle and vascular constructs were bioassembled into a hierarchically organized tissue and co-cultured. By decoupling myogenic and angiogenic differentiation, this strategy overcomes medium incompatibility typical of conventional co-cultures, preserving compartment-specific architecture and function and establishing a versatile platform for muscle-vascular modeling and translational muscle repair.

Curvature provides essential mechanical cues for epithelial cells, playing a key role in cell differentiation and morphology. Repeatable manufacture of precisely controlled curvature in soft hydrogel materials is therefore essential to study epithelial mechanobiology and function. Multiphoton (MP) based biofabrication holds promise due to its high resolution and three-dimensional design flexibility. Here, we leverage MPs advantages while increasing print speed to develop two complementary tools based on replica molding and multiphoton ablation. These can provide scalable hydrogel curvatures with tunable surface properties relevant for epithelial tissue engineering. In replica molding, MP prints are transferred into PDMS used to pattern centimeter scale arrays in hydrogels. In multiphoton ablation, hydrogels are locally degraded to generate precisely controlled curvatures and surface topography. With both methods, we repeatably guide epithelial cells into alveolar and duct-like shapes. Concave alveolar-like surfaces are shown to enhance the formation of thicker epithelial layers. We observe that surface properties, controlled by both tools, could enhance cytoskeletal organization. Using these biofabrication techniques, individual effects of curvature, surface properties, hydrogel composition, and bulk stiffness on epithelial cells can be studied. Both approaches offer high curvature control and throughput, providing a viable alternative to traditional 3D culture and other printing methods.

Understanding how airborne particulates disrupt the alveolar barrier requires in vitro systems that recapitulate both the structure and transport properties of the lung air-blood interface. Here, we report a biodegradable lung alveoli-on-a-chip enabled by porous poly(lactic-co-glycolic acid)/polycaprolactone (PLGA/PCL) membranes with an interconnected porous architecture generated via porogen-assisted phase separation process. The membrane exhibits tunable degradation behavior, allowing progressive increases in surface porosity ([~]40%) and reduction in thickness ([~]3 {micro}m) during culture, while PCL maintains mechanical integrity under dynamic conditions. These degradation-driven structural changes regulate membrane transport properties, leading to enhanced permeability and supporting the formation of a functional epithelial-endothelial barrier under air-liquid interface (ALI) culture with breathing-mimetic cycling strain. Primary human alveolar epithelial and microvascular endothelial cells formed confluent, junctional monolayers on opposing membrane surfaces, exhibiting stable barrier function and high viability throughout the culture period. As a functional application, the platform was used to assess diesel particulate matter (DPM)-induced alveolar injury. Apical exposure to DPM induced dose-dependent cytotoxicity, increased barrier permeability, elevated reactive oxygen species, and DNA damage in both epithelial and endothelial layers, demonstrating trans-barrier propagation of particulate-induced injury. Pharmacological modulation with roflumilast-N-oxide (RNO), a phosphodiesterase-4 (PDE4) inhibitor, selectively attenuated oxidative stress and inflammatory responses, with limited effects on barrier integrity. Together, this work establishes degradable PLGA/PCL membranes as tunable interface materials for lung-on-a-chip systems, where structural evolution during degradation directly governs transport and barrier function. The resulting platform provides a physiologically relevant approach for studying particulate toxicity and therapeutic modulation at the alveolar interface.

The broader clinical application of mRNA therapeutics remains constrained by dose-limiting toxicities, vector-associated immunogenicity, and prolonged tissue retention of lipid nanoparticles (LNPs) in vivo. Here, we report a class of ionizable lipids incorporating a sulfur-bearing hexyl 2-hydroxyethyl sulfide (HHES) motif that decouples mRNA delivery potency from these safety liabilities through dual functionality: the sulfur moiety acts as an intrinsic reactive oxygen species scavenger to suppress oxidative stress, while undergoing oxidative conversion into hydrophilic metabolites to promote rapid systemic clearance. HHES-based LNPs demonstrated a 3.3-fold shorter hepatic half-life and 29-fold lower total hepatic exposure than MC3, while maintaining robust protein expression including functional monoclonal antibody production in vivo. Repeated dosing in non-human primates confirmed negligible systemic, hepatic, or hematological toxicity. Leveraging this safety profile, subretinal HHES LNP delivery achieved up to 57% genome editing efficiency in retinal pigment epithelium, suppressing choroidal neovascularization by [~]65% in a wet age-related macular degeneration model without structural damage or microglial activation. This dual-function design provides a generalizable framework for safe, transient, non-accumulative mRNA nanomedicines.

In mammalian organisms, native tissue function depends on precise spatial organization down to the cellular level. Reconstituting tissue architectures in 2D in vitro platforms can provide a means to study direct and indirect cell-cell interactions in a variety of tissue contexts while remaining compatible with high-throughput assays and high-resolution live imaging. We combine cost-effective stereolithography leveraging 3D printing with replica molding to stencil spatially defined, multicellular culture systems with sub-millimeter resolution onto planar substrates. The system is designed for ease of use, requires no complex fabrication setups and scales readily to 96-well plates. Sequential stencil application and removal under a biosafety cabinet enables controlled positioning of multiple cell types and supported the maturation of tissue assemblies. We demonstrate the utility of this stencil-based patterning strategy in three applications. First, we employ a combination of two circular stencils to recreate a structural feature characteristic for the tumor microenvironment of solid tumors: the encapsulation of colorectal cancer cells by cancer-associated fibroblasts. Resulting cell patternings reproduce native tissue dynamics of the densely packed tumor tissues, in which cancer-associated fibroblast cells actively compress the cancer cells and confer targeted therapy resistance. Second, we probe the synthetic, diffusible morphogen system synNotch in patterned cell patches, where GFP-releasing cells generate a ligand-dependent gradient. Third, we recapitulate the characteristic crypt-villus architecture of the mammalian intestine by patterning intestinal organoids within a stencil-restricted crypt region and allowing differentiating cells to collectively migrate along a designed villus axis. The presented strategy allows for rebuilding multicellular tissue architectures in vitro with biologically relevant spatial precision for high-throughput drug screenings and dissection of tissue-specific cellular interactions.

Engineering three-dimensional neuronal tissues with defined architecture and functional connectivity remains a critical challenge for applications in disease modeling, drug discovery, and regenerative medicine. Recently, a variety of fabrication methods have arisen, such as bioprinting or manual assembly of organoids, but often struggle with scalability, reproducibility, or maintaining cell viability. Here, two scaffold-free acoustic levitation bioreactors are introduced: one optimized for the culture of uniform neuronal spheroids, and another designed for the structuration of assembloids composed of distinct neuronal identities. Using acoustic standing waves, these platforms enable the contactless manipulation of cells and aggregates, facilitating the formation of highly viable functionally mature spheroids. This study shows that both striatal and cortical cell aggregates formed in acoustic levitation self-organize into spheroids within 24 hours and remain viable up to 10 days under these particular culture conditions without medium renewal. These neuro-spheroids demonstrate healthy development with increased growth and typical terminal differentiation and synaptic maturation. Moreover, concentric cortico-striatal assembloids were successfully structured and cultivated using optimized acoustofluidic chips. Offering versatile and scalable tools for engineering complex neuronal networks, acoustic levitation reveals itself as an innovative approach to 3D neuronal tissue modeling, with broad implications for bioengineering, regenerative medicine and fundamental neuroscience research.

Stimulator of interferon genes (STING) is a promising therapeutic target for cancer immunotherapy, but agonists are often rendered ineffective by the loss of STING expression in cancer cells. Here we engineer a multivalent peptide-polymer conjugate material that can easily be delivered to the cytosol, where it mimics key protein interactions from the missing STING protein to directly activate downstream innate immune signaling. While previously developed STING mimicking therapeutics use nearly the full STING protein, this material contains only a 39 amino acid peptide from the STING C-terminal tail that includes interaction motifs for downstream kinase TBK1 and transcription factor IRF3. Conjugation of multiple peptide copies to a negatively charged polymer backbone mimics the multivalent protein-protein interactions of the oligomerized STING signaling complex, activating TBK1 and IRF3 as well as the transcription of downstream genes in both STING-proficient and STING-silenced cancer cell lines. We optimize a lipid nanoparticle formulation to deliver this conjugate material intracellularly, allowing for its application as an immunotherapy for ovarian cancer. Treatment with the STING mimicking conjugate material promoted the production of type I interferons, repolarization of myeloid cells to an anti-tumor phenotype, and recruitment of T cells to tumors in mice. This treatment ultimately led to tumor regression and extended survival in multiple mouse models of metastatic ovarian cancer. Overall, this work highlights the potential of peptide-polymer conjugate mimics of STING to therapeutically activate innate immune signaling.

The cytosolic delivery of therapeutic proteins remains one of the most persistent challenges in modern drug delivery. Here, we report the discovery and characterization of an encapsulin-based protein nanocage, QtEnc, with unexpected permeability properties and the ability to internalize cargo proteins in vitro, fundamentally departing from existing protein nanocage cargo loading paradigms. This permeability enables simple, rapid, and single-step post-assembly cargo loading, accommodating cargos as large as 482 kDa, and allowing multiplexed cargo co-encapsulation with tunable ratios. Leveraging this property, we develop a modular QtEnc-based NanoCarrier (QtEncNC) with a pH-responsive cargo detachment module and an endosomal escape module, enabling low pH-triggered cargo release from assembled shells and subsequent endosomal escape for cytosolic delivery. Using a cytotoxic protein, BLF1, as a proof-of-concept QtEncNC payload, we demonstrate efficient cytosolic protein delivery in HeLa cells. These findings establish QtEncNC as a versatile and modular platform for cytosolic protein delivery with broad biomedical potential.

Size exclusion within biological hydrogels imposes a fundamental constraint on the design of nanocarriers, limiting the transport of cargo-loaded and structurally complex materials through mucus barriers. While surface passivation strategies are commonly used to improve compatibility, they do not address steric limitations imposed by the polymer network. Here, we introduce mechanical flexibility as an independent materials design parameter to expand the functional transport window of nanocarriers in mucus. Using programmable DNA origami to decouple flexibility from size and surface chemistry, we show that increased structural compliance enhances transport under steric confinement by facilitating passage through confined network pores. When surface-driven aggregation dominates, passivation is required to restore transport, after which flexibility provides additional gains. Together, these results establish mechanical flexibility as a general materials design strategy for improving transport under size-constrained conditions, with implications for nanocarrier engineering across biological barriers.

Microneedle (MN) patches have emerged as a highly efficient platform for localized drug delivery, showing great promise in cancer therapy due to their ability to enable precise drug administration. However, conventional MN systems are limited by the low drug-loading capacity of their tips and primarily rely on biologically inert, non-therapeutic matrices for structural support, which restricts further gains in antitumor efficacy. Herein, we present a strategy turning toxicity into therapy by constructing palladium nanoparticle-loaded polyvinyl alcohol/polyethyleneimine (PVA/PEI@Pd) hydrogel microneedles (PPPd-MNs), which exploit the intrinsic cytotoxicity of PEI for synergistic melanoma therapy. The PPPd-MNs efficiently catalyze the deprotection of a doxorubicin prodrug (P-DOX), enabling in situ generation of active doxorubicin (DOX). Notably, the PEI matrix serves a dual function: acting as a robust ligand to stabilize Pd catalysts and functioning as a therapeutic agent that disrupts cancer cell membranes. Both in vitro and in vivo experiments demonstrate that the combination of Pd-mediated bioorthogonal activation of DOX and PEI-induced membrane damage achieves a remarkable synergistic therapeutic outcome in a murine melanoma model, resulting in a tumor inhibition rate of up to 98%. This work repurposes the inherent cytotoxicity of the carrier material as an active therapeutic component, offering a novel paradigm for the design of high-performance bioorthogonal catalytic systems.

A programmable 4-input cascade DNA logic gate utilizing toehold mediated strand displacement (TMSD) was implemented on a 3D printed hybrid paper-polymer vertical flow device (3D HPVF) for on/off sensitive and specific fluorescence detection of platelet derived growth factor BB (PDGF BB). Polypropylene was 3D printed directly on paper and thermally cured to create micro paper analytical devices ({micro}PADs). The 3D HPVF comprised of three layers of {micro}PADs enclosed in a casing that clamped each {micro}PAD securely to ensure seamless and efficient wicking between layers. In the presence of PDGF BB, a partially complementary strand to a PDGF B aptamer (PDGF B Apt), cApt, was liberated from a PDGF B Apt/cApt duplex in solution. The solution was then deposited on the 3D HPVF with a dimeric g-quadruplex hairpin. The 4-nucleotide toehold region on the cApt started the hybridization reaction with the dimeric g-quadruplex hairpin (dGH) opening it up allowing formation of a dimeric g-quadruplex structure that binds with thioflavin T (ThT) with enhanced fluorescence intensity at room temperature. The 3D HPVF exhibits a pico molar range of detection from 10pM to 100pM with a 10pM limit of detection (LOD) for PDGF BB concentrations relevant for pregnant women predisposed to early-onset preeclampsia with clear differentiation when compared to similarly competing analytes PDGF AA and AB.

Hydrogels are increasingly recognized as promising therapeutics for arthritic joints, extending their traditional role as mechanical lubricants to modulators of joint immunity. However, the rational design of these materials remains challenging, with progress largely driven by empirical experimentation. To address this, we curated a comprehensive database of 220 hydrogel formulations from 317 published studies and applied an interpretable machine learning (ML) framework to uncover the relationships between hydrogel design parameters and the arthritis severity score. Using a Random Forest algorithm, our model achieved an external validation accuracy of 0.67 in predicting effective hydrogel therapies for arthritis. Analysis revealed a clear hierarchy of design principles: the choice of functional agent, base polymer, and elastic modulus were the most influential predictors of therapeutic efficacy, with composite agents, protein-based polymers, and softer hydrogels most strongly associated with positive therapeutic outcomes. Mechanistic investigations further demonstrated that successful hydrogels promote an anti-inflammatory M2 macrophage phenotype. Benchmarking against classical statistical methods and a large language model framework showed that our ML approach provided more robust, nuanced insights into complex feature interactions. This data-driven framework offers a generalizable blueprint for the rational design of next-generation immunomodulatory hydrogels, paving the way for more effective arthritis therapies.

Continuous monitoring of protein biomarkers could transform the management of acute and chronic diseases. Despite tremendous potential, wearable health monitors have remained largely limited to metabolites and small molecules. A key challenge is the limited availability of biointerfaces that reversibly track low-abundance proteins in vivo without user intervention. Here, we present the Differential Aptalyzer, a minimally invasive hydrogel microneedle platform for continuous monitoring of proteins in skin interstitial fluid. The platform combines high-affinity antibodies for selective target capture with aptamers for reversible electrochemical signal transduction. When integrated into a differential electrochemical chip and pulse-assisted sensor regeneration, this approach enables continuous monitoring of proteins in a wearable format. Using cardiac troponin I (cTnI) as a clinically-relevant model analyte, Differential Aptalyzer offers a broad dynamic range (0.003-0.640 ng/mL) and strong specificity against interfering proteins. Importantly, this platform reliably tracks both rising and falling exogenous cTnI levels injected into healthy mice, as well as endogenously elevated cTnI in a double-knockout mouse model of coronary artery disease, demonstrating its capability in continuous protein monitoring and identifying coronary artery disease cohorts.