Recent Advances in Two-Dimensional MXene-Based Electrochemical Biosensors for Sweat Analysis

Sweat, a biofluid secreted naturally from the eccrine glands of the human body, is rich in several electrolytes, metabolites, biomolecules, and even xenobiotics that enter the body through other means. Recent studies indicate a high correlation between the analytes’ concentrations in the sweat and the blood, opening up sweat as a medium for disease diagnosis and other general health monitoring applications. However, low concentration of analytes in sweat is a significant limitation, requiring high-performing sensors for this application. Electrochemical sensors, due to their high sensitivity, low cost, and miniaturization, play a crucial role in realizing the potential of sweat as a key sensing medium. MXenes, recently developed anisotropic two-dimensional atomic-layered nanomaterials composed of early transition metal carbides or nitrides, are currently being explored as a material of choice for electrochemical sensors. Their large surface area, tunable electrical properties, excellent mechanical strength, good dispersibility, and biocompatibility make them attractive for bio-electrochemical sensing platforms. This review presents the recent progress made in MXene-based bio-electrochemical sensors such as wearable, implantable, and microfluidic sensors and their applications in disease diagnosis and developing point-of-care sensing platforms. Finally, the paper discusses the challenges and limitations of MXenes as a material of choice in bio-electrochemical sensors and future perspectives on this exciting material for sweat-sensing applications.


Introduction
The current healthcare system is taking a transition from a conventional approach, where patients visit a healthcare professional for any medical aid after developing the disease or its symptoms, towards more sophisticated continuous monitoring of vital biomarkers and then informing patients (sometimes by healthcare professionals remotely) of any deviation from the normal level. This enables early detection of any deviation from the level of the normal biomarker and brings in new possibilities for the prevention of disease that are not possible through the conventional system. This requires highly specific and sensitive biosensing platforms for precisely and continuously monitoring the biomarkers. The electrochemical-based wearable sensors are currently being explored on this line and are found to be an excellent sensing platform to address the above challenges [1][2][3]. Thus, wearable electrochemical biosensors can provide continuous, instantaneous data on patients' physiologies via dynamic, non-invasive measures of biomarkers in biofluids without causing discomfort to the wearing individuals [4][5][6][7][8]. In recent years, the growth of wearable electronic technologies that can precisely quantify vital responses such as body temperature, blood pressure, blood sugar level, and heart rate, aiding in depicting and monitoring the

MXenes-A Material of Choice for Biosensors
In contrast to metal nanoparticles-and graphene derivative-based biosensors, MXenes have gained much attention in developing biosensors due to their excellent biocompatibility and high electrical conductivity properties. A suitable sensor possesses high specificity, sensitivity, LOD, quick response, and a wide operating range. In addition, it also needs to be less expensive not only in lab-scale use, but also while scaling up. MXenes have all/most of these very important properties in the development of biosensors [81][82][83][84].

Synthesis Strategies of MXenes
MXenes, a new class of 2D anisotropic nanomaterial, have significantly improved  [60]. Copyright 2019 Elsevier. Table 1. Key analytes present in human sweat and associated with health issues.

Analytes
Health Condition Refs.

MXenes-A Material of Choice for Biosensors
In contrast to metal nanoparticles-and graphene derivative-based biosensors, MXenes have gained much attention in developing biosensors due to their excellent biocompatibility and high electrical conductivity properties. A suitable sensor possesses high specificity, sensitivity, LOD, quick response, and a wide operating range. In addition, it also needs to be less expensive not only in lab-scale use, but also while scaling up. MXenes have all/most of these very important properties in the development of biosensors [81][82][83][84].

Synthesis Strategies of MXenes
MXenes, a new class of 2D anisotropic nanomaterial, have significantly improved from their first discovery in 2011 (Ti 3 C 2 T x ) by the selective etching of the MAX phase precursor Ti 3 AlC 2 [85]. Typically, MXenes are developed by removing the A layer from their MAX phase via specific etching. The synthetic methods of MXenes can be classified into two types: the top-down approach and the bottom-up approach. The top-down approach involves the direct exfoliation of the A layer, while the bottom-up approach is based on 2D ordered growth from atoms/molecules. Both the methods are discussed in detail in the below sections.

Top-Down Approach
The top-down approach involves the direct exfoliation of the bulk parent MAX phase material with its original integrity retained. These exfoliation processes are typically carried out using chemical and mechanical methods. The top-down approach can be further classified based on the type of precursors used, the delamination process, and the etchants involved in the reaction. The following section describes these methods in more detail.

Based on Precursors
Based on the precursors selected, the preparation strategy of MXenes is classified into MAX-phase and non-MAX-phase methods. In the MAX phase-based preparation method, the A layer is eliminated by using specific selective etchants with optimized concentrations and time intervals, followed by filtration and sonication to obtain fewlayered 2D nanosheets. A typical example of this method is the elimination of the A layer from the MAX phase (Ti 3 AlC 2 ) to form Ti 3 C 2 T x MXene by Naguib et al. (Figure 2a), with hydrofluoric acid (HF) as an etchant in room temperature conditions [85].

Based on Precursors
Based on the precursors selected, the preparation strategy of MXenes is classified into MAX-phase and non-MAX-phase methods. In the MAX phase-based preparation method, the A layer is eliminated by using specific selective etchants with optimized concentrations and time intervals, followed by filtration and sonication to obtain fewlayered 2D nanosheets. A typical example of this method is the elimination of the A layer from the MAX phase (Ti3AlC2) to form Ti3C2TX MXene by Naguib et al. (Figure 2a), with hydrofluoric acid (HF) as an etchant in room temperature conditions [85].
Non-MAX phase-based synthesis was obtained from the selective etching of the gallium (Ga) layer from the Mo2Ga2C with 50% concentrated HF acid to form Mo2CTx MXene [86]. In contrast to the known MAX phase formula, the non-MAX phase method of prepared Mo2CTx MXene consists of two A layers of Ga stacked between the Mo2C layers. To confirm perfectly etched treatment, the X-ray diffraction patterns of Mo2CTx and Mo2Ga2C before and after etching were compared (Figure 2b), in which there was a significant reduction in the peak intensity of Mo2C, which confirms the Ga phase was dissolved during the etching process.

Based on Delamination
In general, synthesizing single-layered MXenes requires further processing steps on the multi-layered MXenes, called delamination. The delamination of multi-layered MXenes can be by the intercalation method or through mechanical exfoliation. It was reported that mechanical exfoliation is ineffective because the interlayer interactions in MXenes produced by this method are highly tacky (the interlayer interaction in the MXene is two-to six-folds stronger than that present in graphite and bulk MoS2 [87]). Moreover, the sonication time also affects the lamellar structure, decreases MXene sheet size, and produces defects in the structure [88][89][90].
In the case of the intercalation method, intercalants are introduced to reduce the interlayer spacing, weaken the interaction between the MXene layers, and facilitate the formation of individual nanosheets. This dramatically improves the surface area and abundant surface terminations directly associated with the MXene's electrically conductive properties. The intercalants widely used for intercalations of the MXene are classified into two types: ionic aqueous solutions that include metal hydroxides or halide salts as aqueous solutions [91,92] and organic intercalants such as dimethyl sulfoxide (DMSO) [93], tetrabutylammonium hydroxide (TBAOH) [94], tetra propylammonium Non-MAX phase-based synthesis was obtained from the selective etching of the gallium (Ga) layer from the Mo 2 Ga 2 C with 50% concentrated HF acid to form Mo 2 CT x MXene [86]. In contrast to the known MAX phase formula, the non-MAX phase method of prepared Mo 2 CT x MXene consists of two A layers of Ga stacked between the Mo 2 C layers. To confirm perfectly etched treatment, the X-ray diffraction patterns of Mo 2 CT x and Mo 2 Ga 2 C before and after etching were compared (Figure 2b), in which there was a significant reduction in the peak intensity of Mo 2 C, which confirms the Ga phase was dissolved during the etching process.

Based on Delamination
In general, synthesizing single-layered MXenes requires further processing steps on the multi-layered MXenes, called delamination. The delamination of multi-layered MXenes can be by the intercalation method or through mechanical exfoliation. It was reported that mechanical exfoliation is ineffective because the interlayer interactions in MXenes produced by this method are highly tacky (the interlayer interaction in the MXene is two-to six-folds stronger than that present in graphite and bulk MoS 2 [87]). Moreover, the sonication time also affects the lamellar structure, decreases MXene sheet size, and produces defects in the structure [88][89][90].
In the case of the intercalation method, intercalants are introduced to reduce the interlayer spacing, weaken the interaction between the MXene layers, and facilitate the formation of individual nanosheets. This dramatically improves the surface area and abundant surface terminations directly associated with the MXene's electrically conductive properties. The intercalants widely used for intercalations of the MXene are classified into two types: ionic aqueous solutions that include metal hydroxides or halide salts as aqueous solutions [91,92] and organic intercalants such as dimethyl sulfoxide (DMSO) [93], tetrabutylammonium hydroxide (TBAOH) [94], tetra propylammonium hydroxide (TPAOH) [95], isopropyl amine [96], n-butyllithium [97], and bovine serum albumin (BSA) [98].

HF etching
In the HF etching-based MXene synthesis method, the layered MAX phase is stirred with HF aqueous solution at room temperature with specified concentration and time.
Here, the multi-layered MXene is produced by an etched A layer from the MAX phase, and M-A bonds are replaced by the weak intercalations of T x termination such as (-OH, -F, =O) on the surface of the multi-layered MXene. In agreement with numerous studies in recent times, several etching parameters such as temperature, time, and concentration of etchant play a deterministic role in the standard of the prepared MXene nanosheets. Kumar et al. demonstrated the effectiveness of Ti 3 C 2 T x MXene etched at elevated temperatures for a binder-free supercapacitor application (Figure 3a-d) [99].

Based on Etchants
HF etching In the HF etching-based MXene synthesis method, the layered MAX phase is stirred with HF aqueous solution at room temperature with specified concentration and time.
Here, the multi-layered MXene is produced by an etched A layer from the MAX phase, and M-A bonds are replaced by the weak intercalations of Tx termination such as (-OH,

Non-HF etching
Even though HF is extensively and effectively utilized to prepare MXenes from the MAX phase, it is highly corrosive and hazardous to humans and the environment [100][101][102]. A trace amount of HF remaining in the MXene sample will harm the composite

Non-HF etching
Even though HF is extensively and effectively utilized to prepare MXenes from the MAX phase, it is highly corrosive and hazardous to humans and the environment [100][101][102]. A trace amount of HF remaining in the MXene sample will harm the composite preparation and biological studies carried out further. Thus, the etching of MXenes by non-HF-based solvents is also extensively studied. The alternative etchants used for etching MXenes are weak acids and environmentally friendly bifluorides such as sodium bifluoride (NaHF 2 ), ammonium bifluoride (NH 4 HF 2 ) , and potassium bifluoride (KHF 2 ). Feng et al. demonstrated the synthesis of Ti 3 C 2 T x MXene from the Ti 3 AlC 2 MAX phase with bi-fluoride in a single-stage process. They found that the NH 4 + , Na + , or K + ions enter the interlayer spacing of the MXene and enlarge the interplanar spacing and delamination efficiency [103].
In comparison to carbide-based MXenes, the Al layer is strongly bonded in the nitridebased MXene. Thus, more energy is required to remove the A layer from the nitride-based MXene. Moreover, the nitride-based MXene is less stable and might be able to dissolve in HF [104,105]. To overcome this issue, molten fluoride is used with the support of hightemperature heating to remove the A layer from the nitride-based MXene. Urbankowski et al. heated the Ti 4 AlN 3 powder at 550 degrees for 30 min under an argon atmosphere with molten fluoride, where the free F − ions were active enough to etch the Al layer from the nitride MXene MAX phases [106]. Studies revealed that the etchants are an essential determinant in the surface termination of the synthesized MXene. The fluorine-based etchants increase the percentage of F content in the surface terminations. Therefore, further modifications are needed for particular applications such as biomedical and sensors [107,108].

Bottom-Up Approach
MXenes can also be synthesized by crystal growth methods using a small organic or inorganic molecule as the precursor. The bottom-up approaches enable precise manipulation of the size of the MXene sheet, geometry, and the surface termination groups in the MXene, which are impossible using top-down approaches [109][110][111][112]. However, compared to the top-down approach of MXene preparation, only a few studies have been performed with the bottom-up approach, perhaps due to the difficulty in the bottom-up approach to build atom-by-atom the MXene's complex structure and multi-component atomic-thick layer. Moreover, the mechanisms of interaction between the layers are still not clear.
Hong et al. demonstrated the preparation of Mo 2 N MXene sheets by the chemical vapor deposition (CVD) method with NH 3 as the source in the temperature range of 1080 • C [112]. Similarly, Ren et al., 2015 developed ultrathin α-Mo 2 CT x crystals with excellent stability and defect free by CVD [113]. Buke et al. reported a detailed study on the effect of temperature, time, and copper layer thickness in the preparation of an MXene by the CVD method. In addition, they also found that Mo 2 C MXene crystals formed on the graphene sheets are thinner than those formed on the copper sheets [114]. In continuation to the CVD method, pulse laser deposition (PLD) and salt template methods were also investigated for the preparation of MXenes through a bottom-up approach [115,116].

Electrical Properties
Similar to the MAX phase, a pristine MXene is all-metallic. Thus, the research has been focused on enhancing the conductivity nature of MXenes by engineering their surface chemistry [140,141]. The prepared MXene using etchants leaves a surface termination group (-OH, -F, =O) which will bond to the metal atom in the MXene. Based on the experimental and theoretical studies, a few reports showed that some termination groups may affect the conductive property of the MXene [142][143][144] and electron mobility [145][146][147]. Yan et al. demonstrated an alkaline-treated MXene for humidity sensors, where they showed similar effects of -OH and -O termination groups in enhancing the electrochemical capacitance and sensor properties [107,148,149]. Pandey et al. investigated the electronic properties of M n C n−1 O 2 MXenes (M = Ti, W, Ta, Hf, Sc Mo, Nb, Cr, Zr, Y, Mn, V) [150], in which they found that oxygen-terminated MXenes are favorable for many applications.

Biocompatibility of MXenes
The principal mechanisms of nanomaterials' toxicity could be one of the following: (1) damage to cells, (2) genotoxicity, (3) slow clearance in the renal pathway and accumulation in the organs, and (4) specific toxicity to the neural and reproductive system. Thus, an extensive biological safety evaluation is required to understand all the possible interactions between the nanomaterials and the physiological systems. MXenes, due to their high surface area and hydrophilicity, provide a suitable matrix for fabricating wearable biosensors [169]. Some preliminary results on MXene cytocompatibility and biocompatibility are already explored in the literature. Ti 3 C 2 MXene is the most commonly investigated system in biomedical applications [166,[170][171][172]. Liu et al. developed Ti 3 C 2 MXene nanosheets for theranostics applications. They revealed that the MXene nanosheets passed in the bloodstream are excreted through human urine via the renal clearance pathway or accumulated in the tumor tissue due to the enhanced permeability and retention effect (EPR). In this study, the Ti 3 C 2 nanosheets' biosafety and biocompatibility are confirmed by the absence of significant weight loss and the absence of necrotic process. Han et al. investigated the in vivo biocompatibility of Ti 3 C 2 MXene by administering the MXene to mice at elevated dosages (6.25, 12.5, 25, and 50 mg/kg), which resulted in excellent in vivo biocompatibility at 25 mg/kg [173]. In a similar study, Zong et al. developed a Ti 3 C 2 -GdW10 nanocomposite for cancer theranostics application. The in vivo biocompatibility studies concluded that up to 50 mg/kg of Ti 3 C 2 -GdW10 nanocomposite was biocompatible using female Kunming mice [174]. Similar results have been reported for systematic cytocompatibility and biocompatibility of tantalum carbide-based MXene Ta 4 C 3 [171,175] and niobium carbide MXene Nb 2 C [176,177].

MXene-Based Electrochemical Sweat Sensors
Due to the fascinating physiochemical properties of MXenes and their composites, they have gained strong attention in biosensing with their excellent sensitivity, specificity, LOD, mechanical properties, and biocompatibility [136,144,178,179].

Glucose Sensing
Peng et al. developed a Pt/Ti 3 C 2 MXene-based wearable, flexible non-enzymatic electrochemical biosensor for continuous monitoring of glucose in sweat [180]. Figure 4a-c illustrates the fabrication process of the sensor, microfluidic patch, and integration of the flexible sensor to the prepared patch. Figure 4d-f shows the conceptual scheme of the sensor for glucose detection, the cross-sectional view of the sensor on the skin, and the oxidation mechanism of the material and analyte. Figure 4g,h shows the electrochemical reaction mechanism of Pt/Ti 3 C 2 coated on a glassy carbon electrode (GCE) and fabricated flexible sensor. This system offers a LOD of 29.15 µmol L -1 with a correlation coefficient of 0.9793.

Glucose Sensing
Peng et al. developed a Pt/Ti3C2 MXene-based wearable, flexible non-enzymatic electrochemical biosensor for continuous monitoring of glucose in sweat [180]. Figure 4ac illustrates the fabrication process of the sensor, microfluidic patch, and integration of the flexible sensor to the prepared patch. Figure 4d-f shows the conceptual scheme of the sensor for glucose detection, the cross-sectional view of the sensor on the skin, and the oxidation mechanism of the material and analyte. Figure 4g,h shows the electrochemical reaction mechanism of Pt/Ti3C2 coated on a glassy carbon electrode (GCE) and fabricated flexible sensor. This system offers a LOD of 29.15 µmol L -1 with a correlation coefficient of 0.9793.  In another work by Park et al., 2022, a butterfly-inspired hybrid epidermal biosensing (bi-HEB) patch, made with carbon Ti 3 C 2 T x MXene-based nanocomposite, was developed for simultaneous monitoring of glucose and electrocardiograms (ECGs) in human subjects while performing indoor physical activities [73]. This system demonstrated excellent sensitivity of 100.85 µAmM −1 cm −2 within physiological levels (0.003−1.5 mM). Moreover, variations in pH and temperature from on-body sweat monitoring are calibrated. for quick, responsive glucose detection. This system has a LOD of 2 µM in a concise time of fewer than 3 s with a sensitivity of 169.49 µA/(mM·cm 2 ) in the range of 2 µM-0.4 mM concentration of glucose [181]. Hosseini et al. fabricated a Ti3C2/nickel-samarium-layered double hydroxide for enzyme-free real-time glucose detection. The glucose sensing was investigated with the DPV method, resulting in a LOD value of 0.24 µM and a linear range from 0.001 to 0.1 mM and 0.25-7.5 Mm [182]. Li et al. demonstrated a highly integrated wearable sensing paper (HIWSP) for electrochemical analysis of glucose and lactate from sweat [183]. This paper composition contains hydrophobic-layered protecting wax, a hydrophilic layer for sweat diffusion, and a Ti3C2Tx MXene/methylene blue composite. This sensor had a sensitivity of 2.4 nA µM −1 and 0.49 µA mM −1 for the simultaneous detection of glucose and lactate, respectively. In another work, Lei et al. fabricated a high-performing wearable biosensor for in vitro perspiration analysis [184]. This multifunctional biosensor is designed by incorporating Ti3C2Tx MXene and Persian blue composite for durable and sensitive monitoring of glucose and lactate in sweat.  Li et al. demonstrated a highly integrated wearable sensing paper (HIWSP) for electrochemical analysis of glucose and lactate from sweat [183]. This paper composition contains hydrophobic-layered protecting wax, a hydrophilic layer for sweat diffusion, and a Ti 3 C 2 T x MXene/methylene blue composite. This sensor had a sensitivity of 2.4 nA µM −1 and 0.49 µA mM −1 for the simultaneous detection of glucose and lactate, respectively. In another work, Lei et al. fabricated a high-performing wearable biosensor for in vitro perspiration analysis [184]. This multifunctional biosensor is designed by incorporating Ti 3 C 2 T x MXene and Persian blue composite for durable and sensitive monitoring of glucose and lactate in sweat.  Ti3C2Tx MXene nanoflakes decorated ZnO tetrapods (ZnO TPs), a graphene oxide composite, and a skin-attachable enzymatic electrothermal glucose sensor was fabricated by Myndrul et al. [186]. This system showed a LOD of 17 µM with a broad linear detection range of 0.05-0.7 mM for detecting glucose from human sweat. Figure 7a-d shows stretchable electrodes at various strains, glucose sensing performance at applied strain (artificial sweat), skin-attachable sensor performance, and current density changes in the fabricated sensors on volunteer sweet consumption for glucose monitoring. Ti 3 C 2 T x MXene nanoflakes decorated ZnO tetrapods (ZnO TPs), a graphene oxide composite, and a skin-attachable enzymatic electrothermal glucose sensor was fabricated by Myndrul et al. [186]. This system showed a LOD of 17 µM with a broad linear detection range of 0.05-0.7 mM for detecting glucose from human sweat. Figure 7a-d shows stretchable electrodes at various strains, glucose sensing performance at applied strain (artificial sweat), skin-attachable sensor performance, and current density changes in the fabricated sensors on volunteer sweet consumption for glucose monitoring.

Cortisol Sensing
Park et al. developed a microfluidic integrated wearable impedimetric immune sensor based on Ti3C2Tx MXene incorporating laser-mediated porous graphene. This immune-sensing patch analyzes cortisol from the sweat by collecting sweat samples at one touch using a microfluidic channel network. Thus, wearable immune-sensing patches exhibit a dynamic range of 0.01-100 nM with a LOD of 88pM [188]. In another work, Laochai et al. fabricated Ti3C2 MXene/AuNPs/L-cysteine composited electrodes for realtime detection of cortisol from sweat [71]. Under optimal conditions, this fabricated sensor shows sensitivity with wide linearity of 5-180 ng mL −1 and a LOD value of 0.54 ng mL −1 .

Cortisol Sensing
Park et al. developed a microfluidic integrated wearable impedimetric immune sensor based on Ti 3 C 2 T x MXene incorporating laser-mediated porous graphene. This immunesensing patch analyzes cortisol from the sweat by collecting sweat samples at one touch using a microfluidic channel network. Thus, wearable immune-sensing patches exhibit a dynamic range of 0.01-100 nM with a LOD of 88pM [188]. In another work, Laochai et al. fabricated Ti 3 C 2 MXene/AuNPs/L-cysteine composited electrodes for real-time detection of cortisol from sweat [71]. Under optimal conditions, this fabricated sensor shows sensitivity with wide linearity of 5-180 ng mL −1 and a LOD value of 0.54 ng mL −1 .  (b) Immobilization process. Reproduced with permission from Ref. [71]. Copyright 2022 Elsevier.

Other Analytes
Zhi et al. developed a bioinspired directional moisture-wicking electronic skin for static health monitoring with the support of triboelectric energy harvesting. The device is made of heterogeneous fibrous polyvinylidene fluoride (PVDF) as a hydrophobic layer and polyacrylonitrile (PAN) as the hydrophilic layer with Ti3C2Tx MXene/CNT conductive ink [189]. Zhang et al. fabricated a wireless, battery-free, wearable, skin-interfaced electrochemical sensing patch for K + monitoring from sweat. They used valinomycin as a K + selective carrier specific to K + , enabling the selective detection of K + by the membrane.

Other Analytes
Zhi et al. developed a bioinspired directional moisture-wicking electronic skin for static health monitoring with the support of triboelectric energy harvesting. The device is made of heterogeneous fibrous polyvinylidene fluoride (PVDF) as a hydrophobic layer and polyacrylonitrile (PAN) as the hydrophilic layer with Ti 3 C 2 T x MXene/CNT conductive ink [189]. Zhang et al. fabricated a wireless, battery-free, wearable, skin-interfaced electrochemical sensing patch for K + monitoring from sweat. They used valinomycin as a K + selective carrier specific to K + , enabling the selective detection of K + by the membrane. Figure 9a-e demonstrates the conceptual operation mechanism of the fabricated device, electrode fabrication, sensor fabrication, and CV analysis of the material, followed by EIS analysis. This fabricated sensor measures the K + ion concentration from human sweat with an excellent sensitivity of 63 mV/dec and a high linear detection range from 1 to 32 mM [77]. In another work, Hui et al. designed a flexible electrochemical heavy metal sensor for the non-invasive detection of Cu and Zn ions in human sweat with a material composition of Ti 3 C 2 T x /multi-walled carbon nanotubes (MWCNTs) in a layer-by-layer self-assembly [187]. The square wave anodic stripping voltammetric method (SWASV) was used for electrochemical analysis. The resulting LOD of Cu and Zn ions is 0.1 and 1.   [190]. In another work, Saleh et al. fabricated inkjet-printed Ti3C2Tx MXene-based electrodes for cutaneous biosensing [74]. They demonstrated an exciting finding on Inkjet-printed MXene electrodes for Na + ion detection and cytokine protein with 3.9 mV per decade by simple changes in the functionalization of the target analyte. These findings simplify the fabrication of wearable electronic platforms that enable multimodal biosensing. Qiao et al. developed a Ti3C2 MXene@TiO2 (anatase/rutile) ternary heterostructured electrode to detect phosphoprotein in sweat. They showed a LOD of 1.52 µM with a broad linear range from 0.01 to 1 mg/mL [191]. Chen et al. developed a  [190]. In another work, Saleh et al. fabricated inkjet-printed Ti 3 C 2 T x MXene-based electrodes for cutaneous biosensing [74]. They demonstrated an exciting finding on Inkjet-printed MXene electrodes for Na + ion detection and cytokine protein with 3.9 mV per decade by simple changes in the functionalization of the target analyte. These findings simplify the fabrication of wearable electronic platforms that enable multimodal biosensing. Qiao et al. developed a Ti 3 C 2 MXene@TiO 2 (anatase/rutile) ternary heterostructured electrode to detect phosphoprotein in sweat. They showed a LOD of 1.52 µM with a broad linear range from 0.01 to 1 mg/mL [191]. Chen et al. developed a flu-oroalkyl functionalized F-Ti 3 C 2 T x /polyaniline (PANI) superhydrophobic skin-attachable and wearable electrochemical pH biosensor for real-time perspiration analysis [192]. Thus, the schematic view of the sensing device setup, pH analysis of sweat in males/females, and comparison of measured values from the fabricated patch with other equipment are discussed in detail in Figure 10a- fluoroalkyl functionalized F-Ti3C2Tx/polyaniline (PANI) superhydrophobic skinattachable and wearable electrochemical pH biosensor for real-time perspiration analysis [192]. Thus, the schematic view of the sensing device setup, pH analysis of sweat in males/females, and comparison of measured values from the fabricated patch with other equipment are discussed in detail in Figure 10a-d.  [192]. Copyright 2022 American Chemical Society.

Summary and Outlook
In recent years, more attention has been paid to MXenes due to their outstanding characteristic features such as high conductivity, surface area, oxidation and redox capacity, biocompatibility, and significant electrocatalytic and electrochemical properties. Moreover, MXenes also hold great potential as conductive substrates for various electrode-based devices, especially for specific-patterned electrodes. This paves the way for the fabrication of highly integrated electrochemical sensing gadgets such as wearable sensors for non-invasive monitoring of biomarkers in the body fluid (sweat) and miniaturized electronic devices for practical standards. However, for the faster translation of MXene-based electrochemical sensors from lab to market, we propose the following research directions in the near future: (i) Device fabrication with precise nano-/micro-patterned structures are essential to enhance the performance of wearable sensors, and their shortcomings cannot be ignored [193].
(ii) It is challenging to synthesize MXenes on a large scale due to their atomic-thick layer design. To overcome this, future research should address the industrial-scale nanostructured design of MXenes and cost-effective technologies [194,195].  [192]. Copyright 2022 American Chemical Society.

Summary and Outlook
In recent years, more attention has been paid to MXenes due to their outstanding characteristic features such as high conductivity, surface area, oxidation and redox capacity, biocompatibility, and significant electrocatalytic and electrochemical properties. Moreover, MXenes also hold great potential as conductive substrates for various electrode-based devices, especially for specific-patterned electrodes. This paves the way for the fabrication of highly integrated electrochemical sensing gadgets such as wearable sensors for noninvasive monitoring of biomarkers in the body fluid (sweat) and miniaturized electronic devices for practical standards. However, for the faster translation of MXene-based electrochemical sensors from lab to market, we propose the following research directions in the near future: (i) Device fabrication with precise nano-/micro-patterned structures are essential to enhance the performance of wearable sensors, and their shortcomings cannot be ignored [193].
(ii) It is challenging to synthesize MXenes on a large scale due to their atomic-thick layer design. To overcome this, future research should address the industrial-scale nanostructured design of MXenes and cost-effective technologies [194,195].
(iii) The malleable structure is crucial in changing the conductivity and internal resistance of the MXene. Hence future research should focus on the enlargement of interlayer distance in MXenes.
(iv) The problems with the flexibility of MXenes can be solved by performing focused research on MXene-polymer nanocomposites using biocompatible polymers [196].
(v) MXene-based biosensors strongly rely on nanohybrid biocompatibility. Thus, there should be focused research on the surface chemistry of MXenes to solve the problems based on the affinity and stability of biomolecules on MXene surfaces [197].
(vi) In the case of wearable sensors, MXene nanomaterial is oxidized when continuously in contact with air. This reduces the conductivity and affects the sensing ability. However, the external polymer coating to prevent oxidation in the MXene affects the breathability and comfort of the wearable biosensors. Thus, an in-depth understanding is needed to design sensors that could maintain the conductivity of the MXene without losing the convenience of the user [169].
Author Contributions: S.G. and A.P. conceptualized the paper; S.G. and K.R. wrote the paper; T.K. and A.P. reviewed and edited the manuscript. All authors have read and agreed to the published version of the manuscript. Data Availability Statement: All the data and materials that support the results or analyses presented in their paper are freely available upon request.

Acknowledgments:
The authors gratefully acknowledge the financial support of this project by the Ministry of Science and Technology, Taiwan (MOST 111-2221-E-027-085). S.G, A.P and K.R. would also like to acknowledge the support received from the Center for Biomaterials, Cellular, and Molecular Theranostics (CBCMT); the School of Advanced Sciences (SAS); and the Periyar Central Library of Vellore Institute of Technology, Vellore.

Conflicts of Interest:
The authors declare no conflict of interest.