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Electrochemical Determination of Kynurenine Pathway Metabolites
Kynurenine pathway (KP) is the major catabolic route of tryptophan, which generates an important enzyme cofactor (NAD+) and a variety of bioactive metabolites (so-called kynurenines) with immunosuppressive functions or neuroprotective, antioxidant, and toxic properties. It is involved in a variety of physiological processes, especially in conditions associated with immune dysfunction, central nervous system disorders, autoimmunity, infection, diabetes, and cancer. In normal conditions, tryptophan depletion via KP is initiated by the liver enzyme tryptophan 2,3-dioxygenase (TDO) and the extrahepatic enzyme - indoleamine 2,3-dioxygenase (IDO) that contributes minimally to this process (5–10%). The extrahepatic KP becomes quantitatively more significant under conditions of immune activation. KP metabolites are frequently found in biofluids, tissues, and cell-delivered material at low nanomolar or low micromolar concentration levels. However, in disease conditions, abnormal tryptophan metabolism can be accompanied by changes in levels of KP metabolites.
1. Kynurenine Pathway
Exploration of the role of tryptophan metabolism provides novel diagnostic and treatment opportunities, however, it requires reliable methods for quantification of its metabolites in a variety of biological samples.
2. Developments in Electrochemical Sensors for the Determination of Kynurenine Pathway Metabolites
|Structure||Eox * [V]
(pH = 7.7)
|Isoelectric Point |
Advantages of the GCE and BDDE surface modification have also been emphasized during the design of voltammetric sensors for Kyn . Electrochemical deposition of Bi film onto the BDDE surface presents an easy and rapid way to improve the sensor sensitivity toward Kyn measured by DPV and it reaches a low LOD (30 nM) . The coating of a GCE surface with a thin layer of Nafion polymer allows for detection of lower contents of Kyn, as the cationic form of this molecule can be pre-concentrated onto the electrode surface before the stripping step . The Nafion layer can be formed by a drop-coating method (without the need for sophisticated apparatus) and easily removed by polishing using alumina slurries. Furthermore, Kyn can be effectively accumulated onto the Nafion-coated GCE at the potential of +0.5 V in 0.1 M H2SO4, before being stripped by scanning potential toward more positive values . This strategy allows for working with diluted samples and decreases some interferences delivered from the sample matrix components. The applicability of Bi film-modified and Nafion-coated sensors was confirmed for the analysis of material derived from cultures of human cancer cells. Karami et al. have also developed the sensor for Kyn quantification in culture medium collected from cancer cells, but applying a multi-stage modification of the surface of the screen-printed gold electrodes (AuSPEs) . The protocol for the modification of AuSPEs’ surface includes the deposition of carboxylated multiwall carbon nanotubes and immobilization of monoclonal antibody (mAb) specific to Kyn.
- Influence of Trp on Kyn and AA signals;
- Overlapping signals from 3HKyn and 3HAA;
- Interferences delivered by tryptophan metabolites formed in other pathways;
- Effect of amino acids;
- Interferences from uric acid, ascorbic acid, and dopamine.
This entry is adapted from 10.3390/s21217152
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