Recognition Properties Research Articles

Synergy of Mutation-Induced Effects in Human Vitamin K Epoxide Reductase: Perspectives and Challenges for Allo-Network Modulator Design.

The human Vitamin K Epoxide Reductase Complex (hVKORC1), a key enzyme transforming vitamin K into the form necessary for blood clotting, requires for its activation the reducing equivalents delivered by its redox partner through thiol-disulfide exchange reactions. The luminal loop (L-loop) is the principal mediator of hVKORC1 activation, and it is a region frequently harbouring numerous missense mutations. Four L-loop hVKORC1 mutants, suggested in vitro as either resistant (A41S, H68Y) or completely inactive (S52W, W59R), were studied in the oxidised state by numerical approaches (in silico). The DYNASOME and POCKETOME of each mutant were characterised and compared to the native protein, recently described as a modular protein composed of the structurally stable transmembrane domain (TMD) and the intrinsically disordered L-loop, exhibiting quasi-independent dynamics. The DYNASOME of mutants revealed that L-loop missense point mutations impact not only its folding and dynamics, but also those of the TMD, highlighting a strong mutation-specific interdependence between these domains. Another consequence of the mutation-induced effects manifests in the global changes (geometric, topological, and probabilistic) of the newly detected cryptic pockets and the alternation of the recognition properties of the L-loop with its redox protein. Based on our results, we postulate that (i) intra-protein allosteric regulation and (ii) the inherent allosteric regulation and cryptic pockets of each mutant depend on its DYNASOME; and (iii) the recognition of the redox protein by hVKORC1 (INTERACTOME) depend on their DYNASOME. This multifaceted description of proteins produces "omics" data sets, crucial for understanding the physiological processes of proteins and the pathologies caused by alteration of the protein properties at various "omics" levels. Additionally, such characterisation opens novel perspectives for the development of "allo-network drugs" essential for the treatment of blood disorders.

Platelet Membrane Biomimetic Manganese Carbonate Nanoparticles Promote Breast Cancer Stem Cell Clearance for Sensitized Radiotherapy.

The presence of cancer stem cells (CSCs) significantly limits the therapeutic efficacy of radiotherapy (RT). Efficient elimination of potential CSCs is crucial for enhancing the effectiveness of RT. In this study, we developed a biomimetic hybrid nano-system (PMC) composed of MnCO3 as the inner core and platelet membrane (PM) as the outer shell. By exploiting the specific recognition properties of membrane surface proteins, PMC enables precise targeting of CSCs. Sonodynamic therapy (SDT) was employed using manganese carbonate nanoparticles (MnCO3 NPs), which generate abundant reactive oxygen species (ROS) upon ultrasound (US) irradiation, thereby impairing CSC self-renewal capacity and eradicating CSCs. Subsequent RT effectively eliminates common tumor cells. Both in vitro cell experiments and in vivo animal studies demonstrate that SDT mediated by PMC synergistically enhances RT to selectively combat CSCs while inhibiting tumor growth without noticeable side effects. Our findings offer novel insights for enhancing the efficacy and safety profiles of RT.

On-column refolding and off-column assembly of parvovirus B19 virus-like particles from bacteria-expressed protein

Virus-like particles (VLPs) are nanometric structures composed of structural components of virions, keeping most of the cellular recognition and internalization properties, but are non-infective as they are deprived of their genetic material. VLPs have been a versatile platform for developing vaccines by carrying their own or heterologous antigenic epitopes. Moreover, VLPs can also be used as nanovessels for encapsulating molecules with therapeutic applications, like enzymes, nucleic acids, and drugs. Parvovirus B19 (B19V) VLPs can be self-assembled in vitro from the denatured major viral particle protein VP2 by equilibrium dialysis. Despite its fair productivity, this process is currently a time-consuming task. Affinity chromatography is used as an efficient step for concentration and purification, but it is only sometimes seen as a method that facilitates the oligomerization of proteins. In this research, we report a novel approach for the in vitro assembly of B19V VLPs through the immobilization of the denatured VP2 into an immobilized metal affinity chromatography (IMAC) column, followed by the on-column folding and the final VLP assembly upon protein elution. This method is suitable for the fast production of B19V VLPs.Key points• Biotechnological applications for inclusion bodies• Efficient single-step purification and immobilization strategies• Rapid VLP assembly strategyGraphical abstract

Two helical Schiff bases with “AIE+ESIPT” characteristics exhibiting selective ion recognition properties

Two racemic Schiff bases (H4L1 and H4L2) were synthesized and characterized, and the crystal structures indicated their helical configurations with equal amounts of P- and M-helicity. The aggregation-induced emission (AIE) and selective ion recognition properties of H4L1 and H4L2 were investigated using UV–vis absorption and fluorescence spectra. The results showed that both H4L1 and H4L2 exhibited obvious “AIE + ESIPT” characteristics in THF/H2O solution upon increasing the water fraction. Both H4L1 and H4L2 displayed an obvious turn-off fluorescence response to Cu2+ and F−, and a slight turn-off response to Fe3+ in DMSO, while they displayed an efficient selective ion recognition of Cu2+ in THF/H2O (1: 1, V/V). The corresponding stoichiometry ratio, association constant, Stern-Volmer quenching constant, and the detection limits were determined via Job's plots, UV–vis spectra titration as well as fluorescence spectra titration experiments. The crystal structures of the complexes obtained from the interactions of H4L1 with Cu2+ and Fe3+ illustrated their binding modes and binding differences, which helped us interpret the ion recognition mechanism of H4L1 and H4L2. In addition, the application of H4L2 on test strips was performed, indicating that H4L2 can be used for the rapid detection of Cu2+ and F− in THF. The acid-responsive fluorescence behavior of H4L2 showed that the white light emission can be obtained upon the addition of hydrochloric acid or aldehyde c into H4L2.

Thermosensitive hydrogel with emodin-loaded triple-targeted nanoparticles for a rectal drug delivery system in the treatment of chronic non-bacterial prostatitis

BackgroundThe complex etiology and pathogenesis underlying Chronic Non-Bacterial Prostatitis (CNP), coupled with the existence of a Blood Prostate Barrier (BPB), contribute to a lack of specificity and poor penetration of most drugs. Emodin (EMO), a potential natural compound for CNP treatment, exhibits commendable anti-inflammatory, anti-oxidant, and anti-fibrosis properties but suffers from the same problems as other drugs.MethodsBy exploiting the recognition properties of lactoferrin (LF) receptors that target intestinal epithelial cells (NCM-460) and prostate epithelial cells (RWPE-1), a pathway is established for the transrectal absorption of EMO to effectively reach the prostate. Additionally, hyaluronic acid (HA) is employed, recognizing CD44 receptors which target macrophages within the inflamed prostate. This interaction facilitates the intraprostatic delivery of EMO, leading to its pronounced anti-inflammatory effects. A thermosensitive hydrogel (CS-Gel) prepared from chitosan (CS) and β-glycerophosphate disodium salt (β-GP) was used for rectal drug delivery with strong adhesion to achieve effective drug retention and sustained slow release. Thus, we developed a triple-targeted nanoparticle (NPs)/thermosensitive hydrogel (Gel) rectal drug delivery system. In this process, LF, with its positive charge, was utilized to load EMO through dialysis, producing LF@EMO-NPs. Subsequently, HA was employed to encapsulate EMO-loaded LF nanoparticles via electrostatic adsorption, yielding HA/LF@EMO-NPs. Finally, HA/LF@EMO-NPs lyophilized powder was added to CS-Gel (HA/LF@EMO-NPs Gel).ResultsCellular assays indicated that NCM-460 and RWPE-1 cells showed high uptake of both LF@EMO-NPs and HA/LF@EMO-NPs, while Raw 264.7 cells exhibited substantial uptake of HA/LF@EMO-NPs. For LPS-induced Raw 264.7 cells, HA/LF@EMO-NPs can reduce the inflammatory responses by modulating TLR4/NF-κB signaling pathways. Tissue imaging corroborated the capacity of HA/LF-modified formulations to breach the BPB, accumulating within the gland's lumen. Animal experiments showed that rectal administration of HA/LF@EMO-NPs Gel significantly reduced inflammatory cytokine expression, oxidative stress levels and fibrosis in the CNP rats, in addition to exerting anti-inflammatory effects by inhibiting the NF-κB signaling pathway without obvious toxicity.ConclusionThis triple-targeted NPs/Gel rectal delivery system with slow-release anti-inflammatory, anti-oxidant, and anti-fibrosis properties shows great potential for the effective treatment of CNP.Graphical

Simple and sensitive sandwich-like voltammetric immunosensing of procalcitonin.

Procalcitonin (PCT) is a reliable biomarker in the early diagnosis of septicemia, pyemia and stroke-associated pneumonia. In this work, through preparing β-cyclodextrin/graphene (CD/GN) nanohybrid as carrier and amplifier simultaneously to band antibodies and probe molecules, a simple and innovative sandwich-like voltammetric immunosensor was proposed for the sensitive and effective determination of PCT. Owing to the host-guest recognition property, the antibodies of PCT can enter into the CD cavities to generate a stable complex; meanwhile, aminopyrene (AP) were introduced as the signal probe and it was adsorbed on the surface of GN via aminopyrine π-πinteraction. Based on the signal change from AP as a response signal which exhibits linearity to the concentration of PCT, a highly sensitive sandwich-type voltammetric immunosensor was developed successfully after optimizing various key parameters. The results demonstrated that the developed sensor had a considerably low detection limit (0.003pgmL-1) and wide linearity of 0.01pgmL-1 to 20.0ngmL-1. This work offered a very simple and sensitive sensing strategy for PCT and other biomarkers via altering the specific antibodies simply, showing great potential applications.

Tetrafluoroterephthalonitrile as an anion‐π donor: theoretical evaluation and application to anion recognition

AbstractThe ability of perfluorinated terephthalonitrile to act as an anion‐π donor fragment in anion receptors is evaluated. New receptors combining an urea and a perfluorinated terephthalonitrile motif into a single architecture have been designed and synthesized. Their molecular recognition properties towards Cl−, Br− and I−, have been studied in solution by means of 1H and 19F NMR as well as photophysical experiments. A complementary electrospray ionization‐tandem mass spectrometry study confirmed the ranking of recognition properties between the receptors. A further theoretical evaluation of binding properties confirmed the association constant trend and suggests a main contribution of the urea motif weakly complemented by a η2‐type anion‐π interaction.

A highly selective and sensitive peptide-based probe for the colorimetric and fluorescent detection of Fe3+, Al3+ and Cr3+ and its application in living cells imaging

A new colorimetric and fluorescent dual-responsive probe named Rb-SSC was designed and synthesized using Rhodamine B and tripeptide (SerSerCys-NH2) as raw reactants. The selective recognition properties in detecting 17 metal ions were investigated by UV–Vis spectroscopy and fluorescence spectroscopy in DMF/water (4:6, v/v, 10.0 mM HEPES, pH = 7.4), and the results showed that Rb-SSC had high selectivity fluorescence response and clear colorimetric analysis for trivalent metal ions (Fe3+, Al3+ and Cr3+). These selectivity were attributed to protonation of Rb-SSC, and the Ksp value of the trivalent ions promotes the formation of hydrolyzed H+. Additionally, the limit of detections (LODs) were found to be 27.4 nM for Fe3+, 28 nM for Al3+ and 62.3 nM for Cr3+, respectively. In addition, MTT assay and cells experiments showed that Rb-SSC has low toxicity and achieved fluorescence imaging of trivalent metal ions in living cells. What's more, the G/B ratio has a good linear relationship with the concentrations of Fe3+ based on the smartphone sensing platform (RGB values), and the limit of detection (LOD) was as low as 0.37 μM.

Generation and characterization of nanobodies targeting human pepsinogens

Human pepsinogens (mainly pepsinogen I and pepsinogen II) are the major inactive precursor forms of the digestive enzyme pepsin which play a crucial role in protein digestion. The levels and ratios of human pepsinogens have demonstrated potential as diagnostic biomarkers for gastrointestinal diseases, particularly gastric cancer. Nanobodies are promising tools for the treatment and diagnosis of diseases, owing to their unique recognition properties. In this study, recombinant human pepsinogens proteins were expressed and purified as immunized antigens. We constructed a VHH phage library and identified several nanobodies via phage display bio-panning. We determined the binding potency and cross-reactivity of these nanobodies. Our study provides technical support for developing immunodiagnostic reagents targeting human pepsinogens.

An ALP-responsive, anionic iridium complex for specific recognition of osteosarcoma cells.

Poor cellular permeability greatly hampers the utilization of anionic Ir(III) complexes, though efficiently emissive and remarkably stable, in cell-based diagnosis. To overcome this barrier, we present the development of an alkaline phosphatase (ALP)-responsive, anionic, and aggregation-induced emission (AIE)-active Ir(III) complex (Ir1) for specific recognition of osteosarcoma cells. Containing phosphate moieties, Ir1 exhibits a net -1 charge, enabling charge repulsion from the cell membrane and resulting in low cellular uptake and good biocompatibility in normal osteoblast cells. Upon ALP-mediated hydrolysis of phosphate groups, the resulting dephosphorylated product, Ir2, demonstrates a positive charge and increased lipophilicity, promoting cellular uptake and activating its AIE properties for specific recognition of osteosarcoma cells that express elevated levels of ALP. This study elucidates the role of ALP as an ideal trigger for enhancing the cellular permeability of phosphate ester-containing Ir(III) complexes, thus expanding the potential of anionic Ir(III) complexes for biomedical applications.

An ECG Deep Learning user identification architecture using ECG sex recognition as a selective parameter

Background:Human user authentication can be implemented by token-, keyword-, or identity-based mechanisms for digital environment session entry (i.e., smartphones, platforms with log-in). Physiological signals, such as ECG, have shown discriminative properties for user identity recognition. Due to ECG hidden nature, it is resilient to public trait exposition, light/noise saturation, or eavesdropping in contrast to fingerprint, facial, voice, or password approaches. ECG might fill those gaps toward a cooperative authentication environment. Methods:This paper proposes a Deep Learning identification scenario in which the inclusion of sex recognition directs the input sample toward a sex-specialized identity classification model, simplifying the discrimination space for each model. The architecture proposed could be suitable for large populations. Our scheme worked with an ECG three-axis pseudo-orthogonal configuration in which each axis is transformed into a time-frequency space. Additionally, we combine each lead matrix in an RGB image, joining the contribution of each wavelet waveform. Results:Our results suggest that it is possible to identify people by using RGB wavelet representations, achieving a classification average of 99.97%. In addition, the inclusion of the sex category for our identification purpose does not significantly affect the classification performance, making it a feasible solution for systems with a larger population. Conclusions:With the features of our database, we have evidence that it is possible to recognize a person’s identity using an ECG sex recognition module through our proposed architecture.

Selective recognition and enrichment of C70 over C60 using an anthracene-based nanotube.

Selective recognition and enrichment of fullerenes (e.g., C60 and C70) remains challenging due to the same diameter and geometrical similarity. Herein, we report a hexagonal anthracene-based nanotube (1) through a one-pot Suzuki-Miyaura cross-coupling reaction. With anthracene-based side walls and pyridine linkers, 1 features a nano-scale tubular cavity measuring 1.2 nm in diameter and 0.9 nm in depth, along with pH-responsive properties. Interestingly, the electron-rich 1 shows high binding affinity (K a ≈ 106 M-1) and selectivity (K s ≈ 140) to C70 over C60 in toluene, resulting from their different contribution of π-π interactions with the host. The protonation of 1 simultaneously alters the electronic properties within the nanotube, resulting in the release of the fullerene guests. Lastly, the selective recognition and pH stimuli-responsive properties of the nanotube have been utilized to enrich C70 from its low-content mixtures of fullerenes in chloroform.

Supramolecular DNA nanogels through host-guest interaction for targeted drug delivery.

DNA hydrogels have been demonstrated with the advantages of good stability, easy modification, and extraordinary biocompatibility, which enables their great application prospects in biosensing, tissue engineering, and biomedicine. Based on the host-guest recognition properties of cucurbit[8]uril (CB[8]), we proposed a general method for constructing functional supramolecular DNA nanogels. Guest molecules have been conjugated into the DNA building units, which could be further crosslinked with CB[8] to construct supramolecular DNA nanogels. At the same time, the aptamer has also been modified into the hydrogel network to achieve cell targeting. These supramolecular DNA nanogels have been demonstrated with a controllable size and multiple stimuli responses, in addition to the excellent biocompatibility, stability and good targeting drug transport ability. Such a host-guest based strategy will provide a molecular library as a "toolbox" for the functionalization of DNA nanogels.

Dissecting the chiral recognition of TLR4/MD2 with Neoseptin-3 enantiomers by molecular dynamics simulations.

Toll-like receptor 4 (TLR4) is a pivotal innate immune recognition receptor that regulates intricate signaling pathways within the immune system. Neoseptin-3 (Neo-3), a recently identified small-molecule agonist for mouse TLR4/MD2, exhibits chiral recognition properties. Specifically, the L-enantiomer of Neo-3 (L-Neo-3) effectively activates the TLR4 signaling pathway, while D-Neo-3 fails to induce TLR4 activation. However, the underlying mechanism by which TLR4 enantioselectively recognizes Neo-3 enantiomers remains poorly understood. In this study, in silico simulations were performed to investigate the mechanism of chiral recognition of Neo-3 enantiomers by TLR4/MD2. Two L-Neo-3 molecules stably resided within the cavity of MD2 as a dimer, and the L-Neo-3 binding stabilized the (TLR4/MD2)2 dimerization state. However, the strong electrostatic repulsion between the hydrogen atoms on the chiral carbon of D-Neo-3 molecules caused the relative positions of two D-Neo-3 molecules to continuously shift during the simulation process, thus preventing the formation of D-Neo-3 dimer as well as their stable interactions with the surrounding residues in (TLR4/MD2)2. Considering that L-Neo-3 could not sustain a stable dimeric state in the bulk aqueous environment, it is unlikely that L-Neo-3 entered the cavity of MD2 as a dimeric unit. Umbrella sampling simulations revealed that the second L-Neo-3 molecule entering the cavity of MD2 exhibited a lower binding energy (-25.75 kcal mol-1) than that of the first L-Neo-3 molecule (-14.31 kcal mol-1). These results imply that two L-Neo-3 molecules enter the cavity of MD2 sequentially, with the binding of the first L-Neo-3 molecule facilitating the entry of the second one. This study dissects the binding process of Neo-3 enantiomers, offering a comprehensive understanding of the atomic-level mechanism underlying TLR4's chiral recognition of Neo-3 molecules.

Dopamine sensing by fluorescent carbon nanoparticles synthesized using artichoke extract.

The practical and easy detection of dopamine levels in human fluids, such as urine and saliva, is of great interest due to the correlation of dopamine concentration with several diseases. In this work, the one-step synthesis of water-soluble carbon nanoparticles (CNPs), starting from artichoke extract, containing catechol groups, for the fluorescence sensing of dopamine is reported. Size, morphology, chemical composition and electronic structure of CNPs were elucidated by DLS, AFM, XPS, FT-IR, EDX and TEM analyses. Their optical properties were then explored by UV-vis and fluorescence measurements in water. The dopamine recognition properties of these CNPs were investigated in water through fluorescence measurements and we observed the progressive enhancement of the CNP emission intensity upon the progressive addition of dopamine, with a binding affinity value of log K = 5.76 and a detection limit of 0.81 nM. Selectivity towards dopamine was tested over other interfering analytes commonly present in human saliva. Finally, in order to perform a solid point of care test, CNPs were adsorbed on a solid support and exposed to different concentrations of dopamine, thus observing a pseudo-linear response, using a smartphone as a detector. Therefore, the detection of dopamine in simulated human saliva was performed with excellent results, in terms of selectivity and a detection limit of 100 pM.

Novel supramolecular deep eutectic solvent (SUPRADES) as a sole chiral selector in capillary electrophoresis for the enantiomeric separation of fluorine-substituted amphetamine analogs

In this study, a novel supramolecular deep eutectic solvent consisting of sulfated-β-CD and citric acid (S-β-CD-CA) is reported for the first time. This innovative system was evaluated as a sole chiral selector in capillary electrophoresis for the enantioseparation of six fluorine-substituted amphetamine analogs, yielding remarkable outcomes. Baseline separations of all amphetamine analogs under study were achieved in less than 21.00 min using the S-β-CD-CA as the chiral selector. It was observed that the addition of 0.050 % v/v S-β-CD-CA into the background electrolyte resulted in the baseline separation of five out of the six fluorine-substituted amphetamine analogs, while in the case of the para-substituted amphetamine analog, 4-fluoramphetamine (4-FA), a higher percentage (0.15 % v/v) was required to achieve baseline enantioseparation. These findings emphasized the potential of this new supramolecular system in providing a class of solvents with promising chiral recognition properties.

A structural decryption of cryptochromes.

Cryptochromes (CRYs), which are signaling proteins related to DNA photolyases, play pivotal roles in sensory responses throughout biology, including growth and development, metabolic regulation, circadian rhythm entrainment and geomagnetic field sensing. This review explores the evolutionary relationships and functional diversity of cryptochromes from the perspective of their molecular structures. In general, CRY biological activities derive from their core structural architecture, which is based on a Photolyase Homology Region (PHR) and a more variable and functionally specific Cryptochrome C-terminal Extension (CCE). The α/β and α-helical domains within the PHR bind FAD, modulate redox reactive residues, accommodate antenna cofactors, recognize small molecules and provide conformationally responsive interaction surfaces for a range of partners. CCEs add structural complexity and divergence, and in doing so, influence photoreceptor reactivity and tailor function. Primary and secondary pockets within the PHR bind myriad moieties and collaborate with the CCEs to tune recognition properties and propagate chemical changes to downstream partners. For some CRYs, changes in homo and hetero-oligomerization couple to light-induced conformational changes, for others, changes in posttranslational modifications couple to cascades of protein interactions with partners and effectors. The structural exploration of cryptochromes underscores how a broad family of signaling proteins with close relationship to light-dependent enzymes achieves a wide range of activities through conservation of key structural and chemical properties upon which function-specific features are elaborated.

A novel approach to chiral separation: thermo-sensitive hydrogel membranes.

The application of membrane technology for separating chiral compounds is hindered due to the restricted availability of chiral recognition sites on the membrane surface. In this study, we propose a novel approach for chiral separation through a selector (bovine serum albumin, BSA) mediated thermo-sensitive membrane system. A thermo-sensitive hydrogel-coated membrane (termed PDTAN) was developed by anchoring poly(N-isopropylacrylamide) (PNIPAm) onto a polyethersulfone (PES) membrane through an adhesive and hydrophilic dopamine hydrochloride (PDA)/tannic acid (TA)/chitosan (Chi) intermediate layer. The results demonstrate outstanding chiral separation efficiency, achieving αL/D = 3.30 for D-phenylalanine (D-Phe) rejection at 40 °C on a BSA-mediated PDTAN membrane system, with significant stability and minimal fouling, surpassing previous findings. Moreover, the PDTAN membrane altered the selective properties of recognition sites in BSA, transitioning from rejecting L-Phe to rejecting D-Phe. Analysis using fourth-order derivative UV-vis, circular dichroism (CD), and in situ Fourier transform infrared spectroscopy (FTIR) techniques revealed a transition in the secondary structure of BSA from α-helix to β-sheet as the temperature increased. This transition, facilitated by hydrogen bonding between BSA and PNIPAm, enabled selective recognition of D-Phe, demonstrating a distinct shift in chiral recognition properties. Importantly, with D-Phe adsorbed onto β-sheet structures of BSA, hydrogen-bond interactions between BSA and the PDTAN membrane were significantly reduced, thereby minimizing membrane fouling and achieving the durability of membrane-based chiral separation.

Temporal integration characteristics of an image defined by binocular disparity cues.

We can correctly recognize the content of an image by presenting all of the elements within a limited time, such as in a slit view or a divided painting image. It is important to clarify how temporally divided information is integrated and perceived to understand the temporal properties of the information-processing mechanism of visual systems. Previous studies related to this topic have often used two-dimensional pictorial stimuli; however, few have considered the temporal integration of binocular disparity for the recognition of objects defined with disparity. In this study, we examined image recognition properties based on the temporal integration of binocular disparity, by comparing that based on the temporal integration of luminance. The effect of element onset asynchrony (the time lag among presented elements) was somewhat similar between disparity and luminance with respect to randomly divided elements. On the other hand, under slit-vision conditions, the tolerance range of spatiotemporal integration for luminance stimuli was much wider than that for disparity stimuli. These results indicate that the temporal integration mechanism in localized areas is common to disparity and luminance, but that for global motion shows differences between the two mechanisms. Thus, we conclude that global motion has little contribution to the temporal integration of binocular disparity information for image recognition.

Intelligent point of care testing for medicine diagnosis

AbstractIntelligent point‐of‐care testing (iPOCT) is a rapidly advancing technology that combines molecular detection and bio‐sensing techniques to achieve instant and accurate analysis through advanced sensors and intelligent algorithms. Molecular detection technologies, such as nucleic acid isothermal amplification, biochip and microfluidics, enable rapid analysis of the chemical composition and concentration of samples, thereby determining their presence. Bio‐sensing technology utilizes the recognition properties of biomolecules in conjunction with sensors to detect specific biomolecules or biological processes. iPOCT technology holds broad prospects for application in the field of healthcare and is expected to enhance detection sensitivity, accuracy, and intelligence through further development. In this review, we delineate the developmental journey of iPOCT, followed by a discussion on the principles of molecular detection and bio‐sensing technology, as well as the design of iPOCT devices for health monitoring and disease diagnosis. Furthermore, we showcased illustrative examples of iPOCT applications in disease diagnosis, highlighting the integration of iPOCT technology with wearable devices and smartphones for attaining more precise and personalized diagnostic results.