Quantification of Glycated Hemoglobin in Total Hemoglobin by a Simultaneous Dual-Signal Acquisition Approach.

The glycated hemoglobin (HbA1c) level, which is defined as the ratio of HbA1c to total hemoglobin (tHb, including glycated and unglycated hemoglobin), is considered one of the preferred indicators for diabetes monitoring. Generally, assessment of the HbA1c level requires separate determination of tHb and HbA1c concentrations after a complex separation step. This undoubtedly increases the cost of the assay, and the loss or degradation of HbA1c during the separation process results in a decrease in the accuracy of the assay. Therefore, this study explored a dual-signal acquisition method for the one-step simultaneous evaluation of tHb and HbA1c. Quantification of tHb: graphene adsorbed carbon quantum dots and methylene blue were utilized as the substrate material and linked to the antibody. tHb was captured on the substrate by the antibody. The unique heme group on tHb catalyzed the production of •OH from H2O2 to degrade methylene blue on the substrate, and a quantitative relationship between the tHb concentration and the methylene blue oxidation current signal was constructed. Quantification of HbA1c: complex labels with HbA1c recognition were made of ZIF-8-ferrocene-gold nanoparticles-mercaptophenylboronic acid. The specific recognition of the boronic acid bond with the unique cis-diol structure of HbA1c establishes a quantitative relationship between the oxidation current of the label-loaded ferrocene and the concentration of HbA1c. Thus, the HbA1c level can be assessed with only one signal readout. The sensor exhibited extensive detection ranges (0.200-600 ng/mL for tHb and 0.100-300 ng/mL for HbA1c) and low detection limits (4.00 × 10-3 ng/mL for tHb and 1.03 × 10-2 ng/mL for HbA1c).

Customized Enhancement of Thermal Sensitivity of Tumors at Different Subcutaneous Depths by Multichannel Lanthanide Nanocomposites.

The photothermal therapeutic effect on tumors located at different subcutaneous depths varies due to the attenuation of light by tissue. Here, based on the wavelength-dependent optical attenuation properties of tissues, the tumor depth is assessed using a multichannel lanthanide nanocomposite. A zeolitic imidazolate framework (ZIF-8)-coated nanocomposite is able to deliver high amounts of the hydrophilic heat shock protein 90 inhibitor epigallocatechin gallate through a hydrogen-bonding network formed by the encapsulated highly polarized polyoxometalate guest. It is superior to both bare and PEGylated ZIF-8 for drug delivery. With the assessment of tumor depth and accumulated amount of nanocomposite by fluorescence, an irradiation prescription can be customized to release sufficient HSP90 inhibitor and generate heat for sensitized photothermal treatment of tumors, which not only ensured therapeutic efficacy but also minimized damage to the surrounding tissues.

A facile electrochemical immunosensor based on EDTA-Pb2+ complexation reaction.

The sensitivity of electrochemical (EC) sensors has been improved through the development of multiple approaches. However, the majority of EC sensors were limited in their practical application by high costs or tedious procedures. Herein, based on ethylenediaminetetraacetic acid (EDTA)-Pb2+ complexation reaction, a facile and affordable immunosensor was designed. Pb2+-magnesium silicate hydrate was served as the sensing substrate. The immunorecognition process was carried out in the Eppendorf tube, and antibody-functionalized Pb2+-polydopamine was utilized as immunoprobe. In the tube, the quantitative and appropriate excess of EDTA was introduced to complex with Pb2+ on the immunoprobes. The remaining EDTA was added to the sensing substrate surface to coordinate with some Pb2+ in it. This leaded to the reduction of the EC signal of Pb2+, which was related to the antigen concentration. Using prostate-specific antigen as the model analyte, the sensitive detection was realized with a low limit of detection (30.49 fg mL-1). Remarkably, the assay results were available within 24 min, sensibly faster than the most existing EC sensors.

Novel dual-responsive hydrogel composed of polyacrylamide/Fe-MOF/zinc finger peptide for construction of electrochemical sensing platform.

Responsive hydrogels have received much attention for improving the detection performance of electrochemical sensors because of their special responsiveness. However, current responsive hydrogels generally suffer from long response times, ranging from tens of minutes to several hours. This situation severely limits the detection performance and practical application of electrochemical sensors. Here, an electrochemical sensing platform was constructed by employing dual-responsive polyacrylamide/zinc finger peptide/Fe-MOF hydrogel (PZFH) as the silent layer, sodium alginate-Ni2+-graphene oxide hydrogel as the signal layer. GOx@ZIF-8, as the immunoprobe, catalyzed glucose to H2O2 and gluconic acid, resulting in the cleavage of immunoprobe as the pH decreased and subsequent release of Zn2+ ions. During the process of Fe-MOF converting from Fe3+ to Fe2+, free radicals were generated and used to destroy the structure of the PZFH. Cysteine and histidine in the zinc finger peptide can specifically bind to Zn2+ to create many pores in PZFH, exposing the signal layer. These synergistic effects rapidly decreased the impedance of PZFH and increased the electrochemical signal of Ni2+. The electrochemical sensing platform was used to detect pro-gastrin-releasing peptide with response times as short as 7 min of PZFH, a wide linear range from 100 ng mL-1 to 100 fg mL-1, and an ultra-low limit of detection of 14.24 fg mL-1 (S/N = 3). This strategy will provide a paradigm for designing electrochemical sensors.

Faraday cage-type self-powered immunosensor based on hybrid enzymatic biofuel cell.

Self-powered immunosensors (SPIs) based on enzymatic biofuel cell (EBFC) have low sensitivity and poor stability due to the high impedance of the immune sandwich and the vulnerability of enzymes to environmental factors. Here, we applied the Faraday cage-type sensing mode on a hybrid biofuel cell (HBFC)-based SPI for the first time, which exhibited high sensitivity and stability. Cytokeratin 19 fragment (CYFRA 21-1) was used as a model analyte. Au nanoparticle-reduced graphene oxide (Au-rGO) composite was used as the supporting matrix for immunoprobe immobilized with detection antibody and glucose dehydrogenase (GDH), also the builder for Faraday cage structure on the bioanode in the presence of antigen. After the combination of immunoprobe, antigen, and the antibody on the bioanode, the Faraday cage was constructed in case the AuNP-rGO was applied as a conductive cage for electron transfer from GDH to the bioanode without passing through the poorly conductive protein. With the assistance of the Faraday cage structure, the impedance of the bioanode decreased significantly from 4000 to 300 Ω, representing a decline of over 90%. The sensitivity of the SPI, defined as the changes of open circuit voltage (OCV) per unit concentration of the CYFRA 21-1, was 68 mV [log (ng mL-1)]-1. In addition, Fe-N-C was used as an inorganic cathode material to replace enzyme for oxygen reduction reaction (ORR), which endowed the sensor with 4-week long-term stability. This work demonstrates a novel sensing platform with high sensitivity and stability, bringing the concept of hybrid biofuel cell-based self-powered sensor.

Anti-Fouling Strategies of Electrochemical Sensors for Tumor Markers.

The early detection and prognosis of cancers require sensitive and accurate detection methods; with developments in medicine, electrochemical biosensors have been developed that can meet these clinical needs. However, the composition of biological samples represented by serum is complex; when substances undergo non-specific adsorption to an electrode and cause fouling, the sensitivity and accuracy of the electrochemical sensor are affected. In order to reduce the effects of fouling on electrochemical sensors, a variety of anti-fouling materials and methods have been developed, and enormous progress has been made over the past few decades. Herein, the recent advances in anti-fouling materials and strategies for using electrochemical sensors for tumor markers are reviewed; we focus on new anti-fouling methods that separate the immunorecognition and signal readout platforms.

Electrochemical sensing interface based on the oriented self-assembly of histidine labeled peptides induced by Ni2+ for protease detection.

To construct an electrochemical sensing interface which was convenient for protease recognition and cleavage, we designed a strategy for directed self-assembly of histidine-tagged peptides on the electrode led by Ni2+ ions for electrochemical detection of prostate specific antigen (PSA). The electrode surface was first functionalized using carboxylated multiwalled carbon nanotubes and then modified with the metal ion chelating agent (5 S)-N-(5-Amino-1-carboxypentyl) iminodiacetic acid (NIA). After the Ni2+ was captured by NIA, the designed immune-functional peptide could be oriented assembly to the electrode interface through the imidazole ring of histidine at the tail, completing the construction of the recognition layer. Therefore, by adding the analyte PSA to identify and shear the immune-functional peptide, the ferrocene in its head was released, resulting in a reduction in the electrical signal, enabling sensitive detection. In addition, the self-assembly layer could be removed by pickling to realize the reconstruction of the recognition layer. Under optimal conditions, the electrochemical sensor had an ultralow detection limit of 11.8 fg mL-1 for PSA, with a wide detection range from 1 pg mL-1 to 100 ng mL-1. In this work, an electrochemical sensing interface based on the histidine-tagged peptide induced by Ni2+ was formed to enable controllable oriented assembly on the electrode surface, and the recognition layer could be reconstructed via pickling, providing a potential approach for the design of repeatable interfaces.

Universal and High-Speed Zeptomolar Protein Serum Assay with Unprecedented Sensitivity.

The accurate detection of trace protein biomarkers is critical for disease diagnosis, healthcare, and pathology research. Currently, the main predicaments of techniques are low sensitivity, prolonged procedures, and the need for specialized devices. Moreover, multistep handling and nonspecific biofouling can lead to high background noise and false positives. To overcome these barriers, a novel ultrasensitive electrochemical platform was developed by combining an electrochemistry approach with the silver mirror reaction to detect proteins at the zeptomolar level. This assay can be accomplished in about only 18 min. As a proof of the concept, human immunoglobulin G (h-IgG) as a model analyte exhibited an ultralow detection limit of 6.31 ag mL-1 (0.04 zeptomoles mL-1). This strategy can be exploited as a universal approach for the ultrasensitive detection of various proteins in clinical diagnostics and point-of-care testing.

Electrochemical Signal Substance for Multiplexed Immunosensing Interface Construction: A Mini Review.

Appropriate labeling method of signal substance is necessary for the construction of multiplexed electrochemical immunosensing interface to enhance the specificity for the diagnosis of cancer. So far, various electrochemical substances, including organic molecules, metal ions, metal nanoparticles, Prussian blue, and other methods for an electrochemical signal generation have been successfully applied in multiplexed biosensor designing. However, few works have been reported on the summary of electrochemical signal substance applied in constructing multiplexed immunosensing interface. Herein, according to the classification of labeled electrochemical signal substance, this review has summarized the recent state-of-art development for the designing of electrochemical immunosensing interface for simultaneous detection of multiple tumor markers. After that, the conclusion and prospects for future applications of electrochemical signal substances in multiplexed immunosensors are also discussed. The current review can provide a comprehensive summary of signal substance selection for workers researched in electrochemical sensors, and further, make contributions for the designing of multiplexed electrochemical immunosensing interface with well signal.

Proton-responsive annunciator based on i-motif DNA structure modified metal organic frameworks for ameliorative construction of electrochemical immunosensing interface.

Reformative exploitation for metal organic frameworks (MOFs) has been a topic subject in electrochemical sensing, in which the loading of electroactive species is always introduced to enable them to generate electrochemical signal. However, insulation shielding of MOFs and flimsy combination method interfere with the signal readout of electroactive dyes when they are co-immobilized on electrode surface, indicating that an amelioration is imperatively proposed to solve these issues. Herein, a proton-activated annunciator for responsive release of methylene blue (MB) based on i-motif DNA structure modified UIO-66-NH2 was presented to design electrochemical immunosensor (Squamous cell carcinoma antigen was used as the model analyte). With the catalysis of a ZIF-8 immunoprobe contained glucose oxidase (GOx) to glucose in test tube, protons are produced in ambient solution and then they can be used as the key to unlock the i-motif functionalized UIO-66-NH2, releasing the loaded MB molecules to be readout on an improved electrode. This stimuli-responsive mode not merely eliminates the insulation effect of MOFs but also provides a firm loading method for electroactive dyes. Under the optimal conditions, the proposed immunoassay for SCCA had displayed excellent performance with a wide linear range from 1 µg mL-1 to 1 pg mL-1 and an ultralow detection limit of 1.504 fg mL-1 (S/N = 3) under the optimal conditions.

Responsive-released strategy based on lead ions-dependent DNAzyme functionalized UIO-66-NH2 for tumor marker.

Signal labeling on electrode interface is an important step during the construction of immunosensor and most signal substances are directly affixed on the immunoprobe or substrate so that some problems such as flimsy labeling method and interference of insulating proteins on electrode surface have been existed to affect their readout. In order to solve above problems in electrochemical immunoassay, a lead ions-decodable autocephalous signal integrator based on UIO-66-NH2 was proposed for the detection of prostate specific antigen (PSA). Briefly, a lead ions-dependent DNAzyme functionalized UIO-66-NH2, in which methylene blue was encapsulated, was independently dispersed in solution phase to be closely associated with the lead sulfide labeled sandwich bioconjugates, and internal methylene blue molecules can be sustained released once a cationic exchange reaction was occurred between lead sulfide label and adscititious silver ions. Based on this designing, immunoassay for PSA was effectively connected with the dynamic behavior of methylene blue molecules through the cleavage of DNAzyme on MOFs surface and performed a wide linear range from 1 pg mL-1 to 10 ng mL-1 and a satisfactory detection limit with 0.34 pg mL-1. The proposed strategy was expected to offer more valuable information for the application of MOFs in early and accurate cancer diagnosis.

In situ growth of electroactive polymers via ATRP to construct a biosensing interface for tumor marker.

A novel biosensing interface for tumor markers was designed based on the atom transfer radical polymerization (ATRP) of poly(isopropenylphenol) (PPPL) in situ initiated by the fixing of p-chloromethyl benzoic acid on the surface of amino-modified electrodes. It was found that the electrochemical activity of PPPL itself can provide sufficient signals for these biosensors, which can avoid signal leakage and streamline the interface modification process. Cu(II) ions absorbed on the carbon spheres and then were released via acid stimulation to act as a catalyst to participate in the interface polymerization with ATRP. As the concentration of targets increased, more Cu(II) ions were released, and the electrochemical signal of polymers was enhanced. Therefore, the sensitive detection of carbohydrate antigen 19-9 (CA19-9) as a model target was achieved, with an ultralow limit of detection of 39 µU mL-1 and wide detection range from 100 µU mL-1 to 100 U mL-1 under optimal conditions. Furthermore, this method achieved satisfying performance in human blood serum with good inter-assay precision (RSD < 6%) and satisfactory recovery of ~ 99-105%. According to the results, this work is of great significance for constructing biosensor interfaces via in situ polymerization. A novel biosensing interface for tumor marker was designed based on atom transfer radical polymerization (ATRP), which poly(isopropenylphenol) with electrochemical signal was fabricated in situ on electrode.

A novel electrochemical sensor based on process-formed laccase-like catalyst to degrade polyhydroquinone for tumor marker.

Methods to improve the sensitivity of electrochemical sensors based on catalytic reactions generally require adscititious or pre-modified catalysts, which make the sensitive detection of sensors extremely challenging. This is because the activity of the catalyst is susceptible to the storage and modification process, such as aggregation during storage or loss of active sites during multi-step modification, which impairs the performance of the sensor. To solve this thorny issue, a novel electrochemical sensor based on a process-formed laccase-like catalyst was constructed for sensitive detection of tumor markers. Cu2+-polydopamine (CuPDA) combined with antibody (Ab2) were employed as copper-containing immunoprobe, which released Cu(Ⅱ) ions under acidic stimulation. Cu(Ⅱ) ions coordinate with the self-assembly cationic diphenylalanine-glutaraldehyde nanospheres (CDPGA) to form a laccase-like catalyst, which had stronger catalytic activity than laccase. The freshly formed catalyst was immediately used to degrade the polyhydroquinone-reduced graphene oxide (PHQ-rGO) composite, resulting in a significant reduction in the current signal. The PHQ-rGO composite plays dual roles of signal substance and substrate on the sensing interface. The proposed electrochemical sensor demonstrated wide linearity for the determination of a model analyte, human epididymis protein 4 (HE4), from 1 pg mL-1 to 100 ng mL-1, and the detection limit was as low as 0.302 pg mL-1 (S/N = 3), which had good consistency with that of electrochemiluminescence method. This process-formed catalyst approach will have potential reference significance for the construction of other sensors.

Fenton reaction-mediated dual-attenuation of signal for ultrasensitive amperometric immunoassay.

In order to alter the complexion of immunoprobe with large impedance as negative factor in sensitivity of amperometric immunosensor, a strategy of Fenton reaction-mediated dual-attenuation of signal was proposed. Herein, metal-polydopamine-Fe3+ composite with the ability of Fenton reaction was initially prepared as immunoprobe for an ultrasensitive immunoassay. The polymerization of dopamine occurred on the surface of ZIF-67 to gain the metal-polydopamine shell, which possessed rich functional groups, negative charge and high specific surface. Then the prepared functional shell was further used to absorb Fe3+ and immobilize labeling antibody as immunoprobe, which was used to construct a sandwich type immunosensor. With addition of H2O2 and aniline, Fenton reaction was triggered to produce hydroxyl radicals, which can not only decrease the current value by degrading methylene blue molecules, but also further initiate aniline to polymerize into non-conductive polyaniline for successive abatement of signal intensity. Therefore, the dual-attenuation of signal model rendered the immunoprobe into a favorable factor and synchronously enhance sensitivity. Expectedly, the detection performance with a linear range from 1.0 × 10-4-100 ng mL-1 and ultralow detection limit of 9.07 × 10-5 ng mL-1 toward neuron-specific enolase was obtained under optimal conditions. This work offered a novel tactic for enhancing sensitivity of immunosensor through the preparation of functional immunoprobe and its rational utilization as signal enhancer.

New immunoprobe: Dual-labeling ZIF-8 embellished with multifunctional bovine serum albumin lamella for electrochemical immunoassay of tumor marker.

A new immunoprobe, which can initiate the sedimentation of Ag nanoparticles (NPs) on an electrode surface, was developed for the electrochemical detection of carbohydrate antigen 72-4 (CA 72-4). To design the immunoprobe, zeolitic imidazolate frameworks (ZIFs) were employed as the carrier to enrich thionine molecules, then bovine serum albumin (BSA) was modified on the electrode surface. Advantageously, BSA, served as an anchor to further attach the labeling antibodies (Ab2) and alkaline phosphatase (ALP) to also be modified on the surface through covalent bonding. To construct the immunosensor, multiwalled carbon nanotube-graphene oxide composites were employed to provide active sites, and the electrodeposited Au NPs were used to immobilize coating antibodies. In the presence of CA 72-4, a sandwich immunosensor was established, and a cascade reaction was initiated to deposit Ag NPs under the catalysis, which can further improve the conductivity of electrode interface. Under the optimal conditions, the immunosensor displayed excellent performance with a wide linear range from 1 µU mL-1 to 10 U mL-1 and an ultralow detection limit of 0.438 µU mL-1 (S/N = 3).

Copper peroxide/ZIF-8 self-producing H2O2 triggered cascade reaction for amperometric immunoassay of carbohydrate antigen 19-9.

Generally, H2O2 is frequently adopted to improve analysis capabilities of various detection systems. However, the addition of H2O2 with relatively higher concentration can lower the bioactivity of antibodies or antigens and the sensing interface stability in most peroxidase and peroxidase-like immunosensors. In order to solve these issues, we designed a novel copper peroxide/ZIF-8 immunoprobe that can self-produce H2O2 to trigger a cascade reaction for the sensitive detection of carbohydrate antigen 19-9. Specifically, CP/ZIF-8 plays a key role as a "signal switch" in the immunosensor. In the presence of HCl, the structures of ZIF-8 and copper peroxide can be broken, producing Cu2+ and H2O2 and a subsequent Fenton-type reaction that generates •OH. The resulting •OH can induce the decomposition of 3-aminobenzeneboronic acid/poly (vinyl alcohol) (PVA) film on the electrode. Although the immunosensor initially showed little current signal due to the poor conductivity pf ZIF-8 and PVA, the current signal was significantly amplified by a HCl-triggered cascade reaction. Under optimal conditions, the immunosensor displayed a wide linear range from 0.0001 to 100 U mL-1 with an ultralow limit of detection of 53.5 µU mL-1 (S/N = 3) for carbohydrate antigen 19-9. Considering these advantages, namely self-producing H2O2 and easy operation, this strategy paves a new way to design other novel sensors.

Ultrasensitive amperometric immunosensor for the prostate specific antigen by exploiting a Fenton reaction induced by a metal-organic framework nanocomposite of type Au/Fe-MOF with peroxidase mimicking activity.

To increase the sensitivity of electrochemical sensor, Fe-MIL-88B-NH2 (Fe-MOF) with peroxidase-like activity is designed for the construction of immunoprobe. The Fe-MOF was prepared by one-step hydrothermalf method using 2-aminoterephthalic acid and iron(III) chloride. For the immunoprobe, it was fabricated by gold nanocomposite/Fe-MOF (Au/Fe-MOF) for the immobilization of labeling antibody (the antibody was used to conjuncting with label materials). The thin layer of Methylene Blue (MB) covered by reduced graphene oxide-gold nanocomposites (Au-rGO) serves as a substrate to covalently fix coating antibodies. The MB as a redox-active species was modified on the glass carbon electrode that can give a strong amperometric signal at 0.18 V (vs. Ag/AgCl). With the participation of H2O2, Fe-MOF can induce the Fenton reaction which degrades MB covered by Au-rGO on the substrate. The rest of MB on the surface of electrode becomes oxidized thereby generating a current signal. Square wave voltammetry (SWV) was used to quantify PSA. Under optimal conditions, the immunoassay is stable, specific and reproducible. It has a lower detection limit of 0.13 pg mL-1 (S/N = 3) and a wide analytical range that extends from 0.001 to 100 ng mL-1. Graphical abstractA sandwich-type amperometric immunoassay based on Fe-MOF-induced Fenton reaction was designed for sensitive determination of prostate specific antigen.

Elevating performance of electrochemical immunosensor via photo-induced microscale hyperthermia in situ.

Catalytic reactions contribute a lot to electrochemical sensing by amplifying electrochemical signals to elevating sensitivity, allowing ultrasensitive sensing of bioindicators. However, the unsatisfactory catalytical performance of catalysts results in low efficiency, limiting its practice in rapid immunosensing. Herein, we demonstrated the potential of photo-induced microscale hyperthermia in accelerating catalysis to enhance sensitivity in the short time. Under near-infrared (NIR) laser irradiation, the period of Fenton-like reaction was significantly reduced, further allowing rapid change of electrical signal on electrode in situ. By constructing a novel immunosensor, efficacious sensing of Squamous Cell Carcinoma Antigen (SCCA) was achieved with improved sensitivity for two times, high timeliness within several minutes and advanced performance of electrochemical immunosensor (linear detection range: 0.1 pg mL-1-1 µg mL-1; limit of detection: 120.2 fg mL-1). To the best of our knowledge, this research is the first work that typifies the photothermal-enhanced catalysis in the electrochemical immunoassays, which illuminates a great direction of developing advanced electrochemical sensing protocol with both favorable capacities and accelerated process.

Label-assisted chemical adsorption triggered conversion of electroactivity of sensing interface to achieve the Ag/AgCl process for ultrasensitive detection of CA 19-9.

Efficient strategies in enhancing sensitivity are pivotal to ultrasensitive detection of tumor markers. In this work, based on the strategy of label-assisted chemical adsorption triggered conversion of electroactivity of sensing interface, a Ag/AgCl process was achieved to enhance sensitivity of the constructed sandwich-type amperometric immunosensor for ultrasensitive detection of carbohydrate antigen 19-9 (CA19-9). Briefly, polydopamine-Ag nanoparticles (PDA-Ag NPs), as signal precursor, combined with labeling antibody were served as labels and graphene oxide-melamine (GO-MA) substrate with chemical absorption capacity was applied as smart sensing interface. After successfully incubating labels, there was primitively no current response due to the poor conductivity between labels and electrode. However, in the presence of H2O2, Ag NPs from labels can be etched into Ag ions, which were adsorbed by GO-MA to form GO-MA-Ag as electroactive substrate. Then, the substrate exhibited a sharp and stable electrochemistry peak of solid-state Ag/AgCl process in the buffer containing KCl. The sensitivity toward detection of CA19-9 was notably enhanced based on the appearance of sharp peak. Under optimum conditions, the designed immunosensor demonstrated a wide working range from 0.0001 to 100 U mL-1 and an ultralow detection limit 0.032 mU mL-1. Thus, utilizing this strategy to construct immunosensor was highly promising in clinical diagnosis for ultrasensitive detection of tumor makers.

Simultaneous multi-signal quantification for highly precise serodiagnosis utilizing a rationally constructed platform.

Serodiagnosis with a single quantification method suffers from high false positive/negative rates. In this study, a three-channel platform with an accessional instrumented system was constructed for simultaneous electrochemical, luminescent, and photothermal quantification of H2S, a bio-indicator for acute pancreatitis (AP) diagnosis. Utilizing the specific reaction between platform and H2S, the three-channel platform showed high sensitivity and selectivity in the biological H2S concentration range. The three-channel platform was also feasible for identifying the difference in the plasma H2S concentrations of AP and normal mice. More importantly, the precision of AP serodiagnosis was significantly improved (>99.0%) using the three-signal method based on the three-channel platform and an optimized threshold, which was clearly higher than that of the single- or two-signal methods (79.5%-94.1%). Our study highlights the importance of constructing a multichannel platform for the simultaneous multi-signal quantification of bio-indicators, and provides rigorous ways to improve the precision of medical serodiagnosis.