Enzyme-Based Electrochemical Biosensor for Therapeutic Drug Monitoring of Anticancer Drug Irinotecan

Abstract

Therapeutic drug monitoring (TDM) is the clinical practice of measuring pharmaceutical drug concentrations in patients' biofluids at designated intervals, thus allowing a close and timely control of their dosage. To date, TDM in oncology can only be performed by trained personnel in centralized laboratories and core facilities employing conventional analytical techniques (e.g., MS). CPT-11 is an antineoplastic drug that inhibits topoisomerase type I, causing cell death, and is widely used in the treatment of colorectal cancer. CPT-11 was also found to directly inhibit acetylcholine esterase (AChE), an enzyme involved in neuromuscular junction. In this work, we describe an enzymatic biosensor, based on AChE and choline oxidase (ChOx), which can quantify CPT-11. ACh (acetylcholine) substrate is converted to choline, which is subsequently metabolized by ChOx to give betaine aldehyde and hydrogen peroxide. The latter one is then oxidized at a suitably polarized platinum electrode, providing a current transient proportional to the amount of ACh. Such an enzymatic process is hampered by CPT-11. The biosensor showed a ∼60% maximal inhibition toward AChE activity in the clinically relevant concentration range 10-10 000 ng/mL of CPT-11 in both simple (phosphate buffer) and complex (fetal bovine serum) matrixes, while its metabolites showed negligible effects. These findings could open new routes toward a real-time TDM in oncology, thus improving the therapeutic treatments and lowering the related costs.

Electrochemical Biosensors Containing Pure Enzymes or Crude Extracts as Enzyme Sources for Pesticides and Phenolic Compounds with Pharmacological Property Detection and Quantification

“…Regarding drug detections, yet Alvau et al [41] proposed a biosensor for therapeutic drug monitoring based on acetylcholinesterase (AChE) and choline oxidase. These are promising biosensors because they also present the possibility of distinct application, for instance, AChE-based biosensors can find application in environmental monitoring, since they can be used for the electrochemical detection of organophosphate and carbamate pesticides.…”

Investigation of Biosensor Potential Component Stability Caused by Influence of External Condition

“…The UV-VIS were examined for the range of 220-350 nm, to observe changes occurring in the peak of approx. 280 nm resulting from the aromatic residues such as tryptophan (Trp), tyrosine (Tyr) and phenylalanine (Phe) present in the protein [25]. An additional experiment was made to verify the stability of Fe 2+ at different concentrations: Fe 2+ (0.5: 1.5: 4 mg/cm 3 ) in time, for samples kept in room conditions.…”

“…Therefore, the recent years show increased interest in development of such devices, both with respect to research and construction. Biosensors are extensively used in:  environment protection [11][12][13][14] where they are a tool to detect water contaminants, harmful gases and various odorants  food industry [15][16][17][18][19][20][21][22] where they are used to detect pesticides examination of product freshness, harmful toxins contained in food, assessment of fungal contamination in grain and detection of specific odorant molecules in nourishment  defense industry, where they are used to detect warfare agents [22]  in medicine, where they are extensively used for diagnostics [23][24][25][26][27][28] to detect markers in cancerous processes, concentration of sugar in blood, detect bacteria and viruses or early identification of diseases, infections and metabolic disorders [29]. Despite the extensive applications of biosensors, there are continuous attempts to improve the existing devices and develop a new generation of sensors.…”

Label-Free Bioelectrochemical Methods for Evaluation of Anticancer Drug Effects at a Molecular Level

“…Indeed, drugs interact with many other different metabolites and cellular components increasing or lowering their concentration. Electrochemistry offers many interesting solutions for the monitoring of these metabolites [84], including fast voltammetry and amperometry at micro-and nanoelectrodes [83,85,86], chip-based electrochemical platforms [87][88][89][90], SECM/SICM configuration for cell imaging and protein, nucleic acid analysis [90,91], electrochemical biosensor technology [92][93][94].…”