3.1. OECT Characteristic Parameters Determination
This section introduces the OECT characteristic parameters that will be used throughout the manuscript to analyze sensing experiments using the CCM and illustrates how these parameters are extracted from continuously recorded transfer curves. The determination of the OECT characteristic parameters will be illustrated from the transfer curves obtained with a PANI channel that was continuously cycled by sweeping the gate potential (V
G) at a low scan rate (
v = 10 mV s
−1) in 1 M HCl. In
Figure 1B, a typical I
DS vs. V
G curve or transfer curve is shown, whereas
Figure 1C shows the first derivative of the transfer curve, which represents the transconductance of the channel [
10].
Three different parameters are usually determined from transfer curves to characterize the OECT response. The first, the threshold voltage (V
TH), represents the minimum gate voltage required to oxidize the polymer (and inject charge carriers). According to the Bernards–Malliaras model for OECT operation, the drain current in the linear regime (low V
DS values) is as follows [
35]:
where W, L, and d, are the channel width, length, and thickness, respectively, μ is the charge carrier mobility, and C* is the volumetric capacitance. Due to the low drain bias used in this work (V
DS = −50 mV), the transistors operate in the linear regime. Consequently, based on the former equation, V
TH can be operationally defined as the gate voltage at which the extrapolated linear fit of I
DS versus V
G intersects I
DS = 0 [
36,
37], as depicted in
Figure 1B. In OECTs, the threshold voltage is often treated as an operational parameter rather than a fixed material constant, as it can be tuned through material design or gate/electrochemical conditions [
36,
38].
On the other hand, the maximum transconductance, g
max, and the gate potential at which the transconductance is maximum, V
G,gmax, can be determined from the first derivative, as shown in
Figure 1C. The latter represents the potential at which the current has the maximum response to changes in the applied gate potential and therefore corresponds to the V
G value at which the OECT signal in terms of I
DS has the greatest sensitivity to changes in the effective gate voltage.
It is common to represent the time evolution of I
DS at a fixed gate voltage; this representation corresponds to the I
DS–
t (drain–source current vs. time) profiles. As shown in this work, time-dependent profiles can be directly measured by recording I
DS at a constant V
G or reconstructed from continuously acquired transfer curves. In both cases, I
DS evolves from an initial steady state value in the (analyte-free) electrolyte, I
DS,0, to a final steady state or plateau value after a perturbation or analyte injection. We define ΔI
corr as the baseline-subtracted change in the drain–source current from the initial condition (
Figure S1). When this parameter is normalized by I
DS,0, the percent change is represented as follows:
The baseline-corrected signal eventually reaches a steady state value, ΔI, which can be quantified as a percent change,
The same procedure employed for the determination of ΔIcorr, ΔI, and ΔI% from IDS was employed for the determination of the analogous parameters ΔVTH,corr, ΔVTH, Δgmax,corr, Δgmax%, and ΔVG,gmax,corr from VTH, gmax, and VG,gmax, respectively.
All the parameters mentioned provide complementary and partially independent descriptors of the OECT response. In the following section, we examine how relying on a single parameter measured at a fixed gate potential can obscure relevant information during sensing experiments, thereby motivating the introduction of the continuous cycling methodology.
First, it is necessary to clarify that transfer curves can be acquired by sweeping the gate potential in either direction, and the resulting responses are not necessarily identical. Owing to the complex processes that couple ionic and electronic charge transport, electroactive polymers often exhibit a certain degree of hysteresis in their redox response. This hysteresis manifests as differences between transfer curves acquired under opposite sweep directions and can also depend on the sweep rate, as will be explored in the following section.
In general, transfer curves can be acquired by sweeping the gate potential in either the turn-off or the turn-on direction of the transistor. In this work, when the gate potential is swept in the positive direction, electrochemical reduction in the polymer is induced, driving it into its insulating state, which is consistent with the OFF state of the transistor. We refer to these transfer curves as “off” curves. Conversely, when transfer curves are acquired while decreasing the gate potential, the polymer is driven into its oxidized, conductive state, and the OECT operates in the ON state. These curves are therefore referred to as “on” curves (
Figure 2A).
3.2. Limitations of Traditional OECT Measurements and Introduction to Continuous Cycling Methodology (CCM)
In this section, we critically analyze the limitations of traditional fixed gate OECT measurements and use representative examples to illustrate why continuous access to the full transfer characteristics is essential for reliable sensing.
When using OECT-based technology for the development of biosensing devices, determining and controlling the characteristic parameters g, V
TH and V
G,gmax, is critical. In particular, the focus is on obtaining transistors with high values of
g (high sensitivity to V
G modulation) [
23,
39] and low values of V
G,gmax and V
TH to avoid polymer degradation or parasitic electrochemical reactions [
40,
41]. To this end, much attention has been paid to the design of channel and gate materials with optimized properties that also allow the incorporation of biomolecules in non-denaturing environments [
10,
42,
43]. Then, the challenge lies in transducing the biochemical events occurring at the channel or gate surface of the OECT into reproducible electrical signals. In many cases, the detection mechanism is based on changes in the interfacial potential due to the adsorption of macromolecules through biorecognition events [
44] or changes in external solution properties such as pH, ionic strength, or counterions flux generated by biochemical reactions [
29]. In all cases, these changes in the physicochemical properties of the gate or channel electrodes lead to changes in the OECT characteristic parameters that can be used as electric signals to monitor sensing events [
45].
However, the evolution in time of these parameters during detection measurements is not commonly used to follow the process in real time. In fact, in many works, the measurement of the transfer curves and the determination of V
TH, V
G,gmax, or
g parameters are evaluated before the sensing event occurs and, in some cases, after analyte addition, but real-time monitoring of these parameters during the interaction of the analyte with the OECT is not usually performed. Instead, the detection signal is obtained by measuring the variation in I
DS at a constant V
G [
24,
46,
47,
48,
49]. Although this methodology has been successfully employed for the detection of several analytes, it misses out relevant information and may present limitations and associated issues. In this section, we will illustrate some of these issues and explore the potential of the continuous cycling methodology proposed here to overcome them by using the response of OECTs based on two of the most widely employed conducting polymers in the construction of electrochemical devices: poly(3,4-ethylenedioxythiophene) (PEDOT) [
50] and polyaniline (PANI) [
27,
51,
52,
53].
In the first place, choosing a constant V
G to perform current monitoring is not trivial. On one hand, the applied V
G must lie within a potential range of high transconductance. However, determining the optimal value of V
G to perform the measurement is not easy since, for transistors based on organic channel materials, the relationship between the output current of the device and the applied potential depends on several factors. For instance, the electrochemical response of polymers may have associated hysteresis, as observed for the case of PEDOT-PAH films in
Figure 2A [
54]. That is, even when sweeping at low rates, the “off” (sweep from −300 to 600 mV) and “on” (sweep from 600 to −300 mV) curves have different responses. For this reason, when applying a constant gate potential, the obtained I
DS will depend on the previous state of the polymer. In this regard, the response of a PEDOT-PAH transistor depends on the gate voltage sweep rate (
v), with the transfer curves shifting towards more positive gate potentials as
v increases, which is characteristic of mixed-conduction polymer systems (
Figure S2). For
v values lower than 20 mV s
−1, the “off” curve (corresponding to the reduction process) shows the same profile independently of the scan rate, meaning that below 20 mV s
−1 the change in the applied potential is slow enough to allow the polymer to reach equilibrium and that no associated diffusion phenomena deform the electrochemical response. However, the “on” curve shifts with scan rate towards lower gate values due to other phenomena that may be associated with restructuring of the polymer chains, generating hysteresis in the current response, even at very low values of
v. As a consequence, the current signal at a given gate potential depends on the initial state of the polymer.
Figure 2B shows the I
DS response of a PEDOT-PAH transistor measured at the same V
G value (V
G = 225 mV), obtained from different initial states of the polymer: off-limit (fully reduced), on-limit (fully oxidized), and open circuit potential (OCP). It can be observed that the changes in the output current depends on the initial state of the polymer, even when they were measured at the same constant gate potential.
This effect is even more notorious for PANI-based OECTs, for which the hysteresis in the response yields a difference of 150 mV between the “on” and “off” curves even at very low scan rates, such as 2 mV s
−1 (
Figure 2C). In this case, the initial state of the polymer has a very important role in the current response of the transistor. In
Figure 2D, I
DS measured at a constant V
G is shown, where differences in output current depending on the initial state of the polymer can be seen. In this experiment, initial gate potentials of 100 mV or −450 mV were applied for 3 min, to fully reduce or oxidize the polymer, respectively. Next, V
G = −250 mV was set, obtaining an output current of −0.57 mA when the initial potential was 100 mV and −0.90 mA, when the initial potential was −450 mV. This means that if the polymer is initially oxidized, applying a constant value of −250 mV yields the polymer in a completely “on” state with low transconductance. Therefore, a sensing event occurring in the gate electrode that generates a shift in the transfer curve will not have a significant effect on the output current. On the contrary, if the polymer is initially reduced, applying V
G of −250 mV will place the polymer in a state of high transconductance, yielding high sensitivity towards analyte detection. A similar effect, but in the opposite direction, occurs at V
G = −150 mV. When the polymer is initially oxidized, applying V
G of −150 mV generates a state of high transconductance in the polymer (I
DS = −0.3 mA), whereas if the polymer is initially reduced, the current is practically null and a sensing event will not be detected. Then, to choose a proper potential to monitor a sensing event, a very thorough study of the polymer behavior and precise control of its redox state before initiating the measurement is required, and even then, any change in the external media or modification of the polymer surface may change the result.
To show how this may affect a sensing experiment, we monitored the change in the signal of a PANI-based OECT upon the addition of NaOH. Changes in the external pH generate changes in the current response of the OECT, since the electrochemical response of PANI strongly depends on the pH. This is the case for many organic composites employed as channel materials in the construction of OECTs and is often harnessed to monitor sensing events involving enzymatic reactions that generate acidic or basic products [
29,
30]. In
Figure 2E, the I
DS vs. V
G profiles of a PANI-based OECT are shown before (blue) and after (purple) the addition of NaOH. A shift in the transfer curves to higher V
G and a decrease in the maximum current, I
DS,max, can be observed. Next, we chose a V
G potential in the high transconductance region to monitor changes in solution pH using the traditional chronoamperometric methodology and to evaluate the influence of the initial state of the polymer on the resulting signal. To this end, initial potentials of 100 mV or −450 mV were applied for 3 min and then changed to −250 mV while I
DS was recorded. Next, NaOH solution was injected to change the solution pH (25 µL of a 20% p/V NaOH were added to a total cell volume of 600 µL of 1 M HCl solution). In
Figure 2F, a marked increase in the current can be observed when NaOH solution was added, as a consequence of the shift in the transfer curve to higher V
G values, when the polymer was initially fully reduced. However, for the experiment performed with the polymer initially oxidized, no appreciable change in the current was observed with the addition of NaOH. The same experiment was performed at −150 mV, where the opposite behavior was found: when the initial state of the polymer was fully oxidized, a marked change in the current was observed upon NaOH solution addition. On the contrary, if the initial state was fully reduced, a very small current was observed, and no changes due to pH change were detected (
Figure S3). These results show the complexity of selecting an appropriate V
G value to perform a detection measurement and the different variables that may affect the result.
In this context, the CCM proposed here allows this issue to be overcome while also providing more information about the processes occurring in the OECT. In this method, the entire transfer curves are continuously recorded during the sensing experiment; therefore, changes in I
DS are registered at all V
G values within the potential range of the transfer curve. Then, the I
DS as a function of time profiles can be reconstructed at each V
G, allowing selection of the value that yields the greatest amplitude of current change from both the “on” and “off” curves (
Figure S4). In
Figure 2G,H, the changes in current, ΔI
corr% and ΔI%, respectively, are shown for the “on” curves (“off” curves are shown in
Figure S5; details of the baseline subtraction procedure in
Section S1). It can be observed that the amplitude of the current change upon the addition of NaOH depends on the V
G analyzed. Then, the potential value that has the highest sensitivity can be selected. In addition, the evolution in time of other parameters can be obtained from the shifts in the transfer curves and their derivatives (
Figures S6 and S7).
Figure 2I shows the changes in V
TH and g
max, demonstrating that these parameters can also be employed to monitor a sensing event. Moreover, reconstruction of the I
DS vs. time profiles from the “off” curves shows that the change in current has different behaviors depending on the potential region analyzed (
Figure S5). For the potential range V
G > −150 mV, there is an increase in current (in absolute value) with the addition of NaOH, while for V
G values below −200 mV, a decrease in signal is observed. This can be explained by taking into account two features of the electrochemical response of the polymer. On one hand, there is a well-known change in the PANI redox potential with increasing pH [
55], which leads to a shift in the transfer curves to higher gate potential values. On the other hand, as pH increases, there is a decrease in PANI conductivity, resulting in reduced transconductance and maximum current values. Because of these coupled phenomena, there is an intersection of the transfer curves that yields different behaviors of the current depending on the potential region analyzed. These results emphasize the relevance of having access to the entire transfer curves and of monitoring the evolution of different parameters to understand the phenomena causing the signal in the transistor. Accordingly, direct quantitative benchmarking against state-of-the-art OECT sensors operating at fixed gate potentials is not straightforward, as the CCM fundamentally changes the way sensitivity and response metrics are defined and extracted.
Another problem that arises from measuring at a constant VG value is that polarization of the channel and gate electrodes can lead to parasitic reactions, generating a component associated with these processes in the OECT signal. As a result, the time required to achieve the stabilization of the device response, as well as the presence of drift in the transistor signal, depends on the VG employed.
As an example, the evolution of the signal of a PEDOT-PAH transistor during the first 20 min after applying a constant V
G depends on the applied potential (
Figure S8). However, sweeping the gate potential between the on and off states avoids polarization, thereby preventing parasitic reactions that add non-desirable components into the device signal. Then, the time required to stabilize the output signal does not vary significantly with the V
G value chosen to monitor the evolution of the current; that is, both the time required to reach stabilization and the drift in the signal are independent of the chosen potential.
Moreover, analysis of other parameters, such as the threshold voltage, can help elucidate the origin of the drift in a measurement and, in some cases, mitigate its effect. For instance, drifts often originate from polymer degradation processes or their detachment from the substrate. In such cases, the OECT current progressively decreases, but separating this contribution from current changes due to an actual sensing event can be a challenge. In this regard, monitoring the threshold voltage can be more appropriate (
Figure S8). In the following sections, we show how this strategy can be harnessed to detect the presence of an analyte through monitoring V
TH under conditions in which the current does not show a clear response.
Taken together, these results demonstrate the relevance of determining different OECT parameters and the potential of monitoring them to improve the performance of OECTs in biosensing measurements. We believe that having all this data available for each experiment is a key factor towards improving reproducibility and achieving a better understanding of the physicochemical processes that take place during detection events. In the following sections, we will show examples of the application of CCM for the detection of different analytes, ranging from monitoring the adsorption of polyelectrolytes and enzymes on polymeric substrates to the biosensing of glucose and urea through biocatalytic reactions.
3.3. CCM Applied to Polyelectrolyte Adsorption Monitoring with OECTs
We now apply the CCM to the monitoring of polyelectrolyte adsorption as a first application example, using this well-characterized system to illustrate how different OECT parameters capture complementary aspects of the adsorption process. In this regard, we monitor the adsorption of poly-styrenesulfonate (PSS), used as a model polyanion, on the PEDOT-PAH channel surface through analysis of the different parameters mentioned above. Adsorption of this macromolecule onto the PEDOT-PAH surface has been previously studied and is driven by electrostatic interactions between negatively charged PSS and positively charged channel surface endowed by the amino groups of polyallylamine [
22,
44].
To this end, the OECT I
DS response was recorded while the gate potential was swept between −200 mV and 600 mV in 0.1 M KCl at 20 mV s
−1 under flow conditions. Then, a 1 mg mL
−1 PSS solution in 0.1 M KCl was injected for 40 min and then rinsed with KCl.
Figure 3A shows the change in transfer response for the “off” curves (on curves are shown in
Figure S9). A shift in the curves to higher gate values is observed. This effect is also evident in
Figure 3B, which shows the transconductance vs. V
G for the transfer curves in
Figure 3A. Then, the V
TH was calculated from each curve and plotted as a function vs. time, as can be seen in
Figure 3C. A rapid increase in V
TH is observed as the charged macromolecule is deposited. The increase in the gate potential required to oxidize the polymer when PSS is adsorbed is related to the incorporation of a negative dopant into the PEDOT-PAH matrix, which stabilizes the positive carriers of the polymer. This effect can also be monitored by following the increase in V
G,gmax (
Figure S10). In addition,
Figure 3D shows g
max values as a function of time, revealing a decrease in this parameter as PSS is deposited on the surface. Next, we continued with the reconstruction of I
DS vs. time profiles usually used to monitor these processes at different constant V
G values. In this case, the I
DS time evolution at different potentials also depends on the selected V
G (
Figure S11). However, some interesting information arises from the comparison of current changes across different V
G.
Figure 3E shows the changes in current at each V
G value obtained by subtracting a linear baseline corresponding to the current before the injection, ΔI
corr, and
Figure 3F shows the total current change after returning to the KCl solution, which is the typically used analytical parameter in OECTs (these results were obtained using the “off” curves; the ones obtained with the “on” curves are shown in
Figure S12). The maximum amplitude of the change in the signal is observed at the potential corresponding to the maximum transconductance of the polymer (380 mV). However, it is interesting to note that a different result is obtained when analyzing the relative changes in the current or normalized current response, ΔI%, which is the ratio between the current change upon analyte addition and the initial current value before the injection. This parameter is often employed to compare the performance of different transistors [
47] and does not necessarily have the greatest amplitude in the V
G,gmax. In fact, in the case of PSS adsorption on the PEDOT-PAH-based OECT, the magnitude of the relative current changes increases with V
G, reaching values of 66% at V
G = 660 mV, compared with a 24% change at V
G,gmax (
Figure S12). These differences depend on each specific system and are not easy to predict; therefore, having all data available allows the selection of the optimal potential to perform the analysis, obtain the signal with the greatest amplitude, and have fair comparison between different devices.
In addition, we compared the OECT response obtained with the CCM with SPR results to evaluate the influence of the methodology in the kinetic profile of the reconstructed curves. To this end, a PEDOT-PAH modified Au-SPR substrate was employed to follow changes in the sensorgram during the injection of 0.1 mg mL
−1 PSS in 0.1 M KCl solution (the same conditions as those employed for the OECT measurements). The kinetic profiles, in terms of both the relative change in the threshold potential and I
DS at V
G = 380 mV, correlate well with the SPR temporal response (
Figure S13), which proves that the CCM allows precise time monitoring of the processes.
3.4. CCM Applied to Enzyme Adsorption Monitoring with OECTs
In this section, enzyme adsorption is investigated using urease and glucose oxidase as representative and practically relevant model systems. Urea and glucose were selected as clinically relevant biomarkers widely used in biomedical diagnostics, while urease and glucose oxidase are well-established enzymes in OECT-based sensing platforms [
29,
30,
44]. In this context, the continuous cycling methodology (CCM) enables the monitoring of both enzyme immobilization and subsequent catalytic activity through the simultaneous analysis of multiple OECT parameters. The anchoring of urease and glucose oxidase on PEDOT-PAH OECTs has been previously studied [
29,
30,
44]. These enzymes are negatively charged in aqueous media at pH 7.4; therefore, due to electrostatic interactions, they are adsorbed onto the protonated polymeric surface. In previous works, we have shown the capability of OECTs to monitor protein deposition on PEDOT-PAH films through changes in the current signal of the device. Here, we report monitoring of the adsorption of urease and glucose oxidase on the OECT channel by following the time evolution of V
TH, V
G,gmax, and g
max, showing the advantages of this analysis methodology.
Integration of urease onto PEDOT-PAH films was performed similarly to polyelectrolyte adsorption. Briefly, under flow conditions, a solution of 10 mM KCl was injected, and I
DS was recorded while the potential was swept between 0 and 800 mV at 10 mV s
−1. Then, 1 mg mL
−1 urease in 10 mM KCl was injected for 32 min, followed by rinsing with 10 mM KCl.
Figure 4 shows the time evolution of g
max, V
TH, and V
G,gmax. These parameters were obtained from both “on” and “off” transfer curves. Transfer curves recorded before and after enzyme deposition show a clear shift toward lower gate potentials (
Figure S14). This variation is also reflected in the time evolution of g
max, V
TH, and V
G,gmax, as shown in
Figure 4A–C. In previous works, such behavior has been attributed to changes in impedance at the polymer/electrolyte interface due to enzyme attachment on the channel surface [
30]. Overall, it is interesting to note that all studied parameters are suitable for monitoring the enzyme integration process.
A similar analysis was performed using glucose oxidase (GOx) instead of urease. In this case, the enzyme injection lasted 29 min, and the behavior of the parameters was opposite to that observed for urease incorporation. In particular, the increase in the conductivity of the channel due to the anchoring of GOx was attributed to stabilization of the positive charge carriers of the polymer by the negative charges of the enzyme, similarly to the behavior observed for PSS adsorption [
44], corroborating the capability of this strategy for monitoring enzyme integration (
Figure S15).
Likewise, a similar approach to that shown above for PSS was applied to compare the OECT response with the SPR signal during urease deposition. In this case, a PEDOT-PAH-modified Au-SPR substrate was employed to follow changes in the sensorgram during injection of 1 mg mL
−1 of the enzyme in 10 mM KCl solution. In all cases, the reconstructed kinetic profiles in terms of V
TH, V
G,gmax, and g
max exhibited a notable correspondence with the SPR sensorgram (
Figure S14), verifying the capability of the CCM to monitor adsorption processes over time with high precision.
3.5. CCM Applied to the Biosensing of Catalytic Reaction with OECTs
Having established the applicability of the CCM to adsorption monitoring, we next demonstrate its use in enzyme-mediated biosensing, where chemical reactions at the channel surface dynamically modify the OECT response. Urease catalyzes the hydrolysis of urea into ammonia and carbon dioxide, yielding an increase in local pH. The catalytic response of enzyme-adsorbed PEDOT-PAH OECTs is shown in this section. For urea detection, the urease-modified OECT mentioned above was exposed to increasing concentrations of urea, ranging from 100 µM to 5 mM in 10 mM KCl at pH 7.4 under flow conditions. Transfer curves recorded between 0 and 850 mV at 10 mV s
−1 exhibited a shift toward higher gate potentials (
Figure S16). As a result, there is an increase in V
TH and V
G,gmax parameters (
Figure 5A,B). In previous works, it was stated that the shift toward higher gate potentials upon pH increase arises from the deprotonation of PAH amino groups, leading to stabilization of PEDOT channel positive charge carriers [
30]. Moreover, the process can be monitored as an increase in g
max, as shown in
Figure 5C.
Figure 5D presents V
TH and V
G,gmax as functions of urea concentration, showing increases in these parameters throughout the entire concentration range studied and a good match between the profiles, suggesting that both parameters can be equally employed for urea detection. A similar behavior was observed for ΔI% and Δg
max% parameters (
Figure 5E), which represent the relative changes in I
DS and g
max, respectively.
As observed for PSS, the time evolution of I
DS at different potentials shows that the device output signal depends on the selected V
G (
Figure S17). From the analysis of ΔI
corr obtained at different V
G values, it can be seen that the maximum signal amplitude occurs at the gate potential corresponding to the maximum transconductance of the polymer (306 mV) (
Figure 5F). On the other hand, relative changes in the normalized current response ΔI
DS % increases with V
G, in a similar fashion to that observed during the monitoring of macromolecule adsorption. These results reinforce the importance of considering the complete set of experimental data in order to identify the gate potential that maximizes signal amplitude and ensures consistent and reliable comparison between devices. Unlike conventional fixed-bias OECT measurements, which typically yield a single sensitivity value, the CCM provides a multidimensional description of device response, allowing sensitivity to be optimized a posteriori by selecting the most informative parameters and gate potentials for a given system.
Furthermore, we have also evaluated the enzymatic detection of glucose through the different parameters with a PEDOT-PAH OECT modified with GOx, showing that the CMM can be successfully employed for monitoring this analyte (
Figure S18). However, some noteworthy results emerged from an experiment performed with a modified electrode channel that suffered degradation, losing polymeric material during the sensing measurement and therefore decreasing the current signal. In previous works, it was reported that I
DS,max is related to polymer thickness [
44]. Therefore, in CMM, the decrease in the maximum current during transfer curve acquisition can be attributed to polymeric material degradation. In general, a stable I
DS is expected during electrolyte monitoring. However, as the OECT degrades, such stability cannot be achieved.
When this kind of issue appears in a sensing measurement, the I
DS shift cannot be clearly associated with either the degradation or the molecular reaction that occurs at the polymeric surface. In the case of glucose sensing, its decomposition into gluconic acid and hydrogen peroxide in the presence of GOx results in a decrease in current, behavior attributed to medium acidification. Therefore, in this case, both the sensing event and the degradation process yield a decrease in the current, and separating one contribution from the other is not possible using only the current signal. In this context, a possible solution is to use parameters that do not strongly depend on the polymer mass to follow the enzymatic reaction.
Figure 6 shows the behavior of a PEDOT-PAH OECT that is degrading while glucose detection is being monitored (glucose was injected from 0.5 to 10 mM in 10 mM KCl at pH 7.4). The evolution of the transfer curves (
Figure 6A) shows a decrease in I
DSmax as the glucose concentration increases, which involves an obstacle for monitoring glucose detection by recording I
DS at a constant V
G of 300 mV (
Figure 6B). Before glucose injection, the I
DS decreases over time, and after glucose injection, a stepwise current shift to lower I
DS values is observed as the concentration increases up to 5 mM. Then, the I
DS continues to decrease; however, no stepwise response is observed upon further increases in concentration. In fact, for values higher than 5 mM, it is not possible to evidence a correlation between current and concentration, and it seems that enzyme saturation has been reached. Otherwise, if we use V
TH to follow the glucose detection (
Figure 6C), it is possible to sense concentrations beyond 5 mM. This issue can be corroborated by applying baseline subtraction to V
TH, yielding a steady state corrected shift (ΔV
TH,corr), and to I
DS, from which ΔI% was computed. The comparison between the percentage change in current and the change in V
TH as function of concentration is shown in
Figure 6D. While ΔI% appears to reach a plateau at 5 mM of glucose, ΔV
TH,corr continues to increase up to 10 mM glucose.
In order to quantitatively compare CCM with traditional I
DS at constant V
G measurements, we performed a glucose sensing experiment using the same transistor under same experimental conditions and applying both methodologies. Specifically, the response of a GOx-modified PEDOT–PAH OECT upon the injection of 5 mM glucose in a 10 mM KCl solution was first recorded by monitoring changes in I
DS at V
G = V
G,gmax, which is presumably close to the most sensitive operating potential. The experiment was then repeated using CCM, allowing the reconstruction of the I
DS response at different V
G values. An increase in signal amplitude is observed when using CCM at the same gate potential employed in the traditional fixed V
G measurement (
Figure S19). Moreover, CCM enables the monitoring of other gate–voltage regions that exhibit even larger response amplitudes.
Finally, the repeatability analysis performed for representative sensing experiments further supports the robustness and versatility of the CCM. Repeated measurements carried out at fixed analyte concentrations (glucose and urea) show stable and reproducible responses, with low dispersion in the extracted parameters, confirming that continuous cycling of the gate potential does not introduce detrimental effects on device performance (
Figures S20 and S21). On the contrary, CCM enables reliable monitoring of sensing events over successive experiments. Importantly, the availability of the complete set of reconstructed transfer curves allows the gate potential at which the current response is maximized to be selected a posteriori, thereby improving the analytical sensitivity without requiring changes in device architecture or measurement conditions. These results highlight the potential of CCM as a general and robust strategy for OECT-based sensing, providing enhanced sensitivity, reproducibility, and interpretability of the electrical signals. Additionally, in the next section, we address how the methodology can be adapted to monitor faster processes by tailoring the cycling protocol.