Development and optimization of a novel conductometric bi-enzyme biosensor for l-arginine determination

• Published in: Talanta (Oxford) Vol. 92; no. 7; pp. 58 - 64
• Main Authors: Saiapina, O.Y., Dzyadevych, S.V., Jaffrezic-Renault, N., Soldatkin, O.P.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.04.2012; Elsevier
• Subjects: Analytical chemistry; Animals; Arginase; Arginase - chemistry; Arginine - analysis; Biological and medical sciences; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Biosensors; Biotechnology; Cattle; Chemical Sciences; Chemistry; Conductometric biosensor; Conductometry; Cross-Linking Reagents - chemistry; Electrochemical methods; Electrodes; Enzymes, Immobilized - chemistry; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; General, instrumentation; Glutaral - chemistry; Hydrogen-Ion Concentration; l-Arginine determination; Limit of Detection; Membranes, Artificial; Methods. Procedures. Technologies; Quality-control analysis; Serum Albumin, Bovine - chemistry; Urease; Urease - chemistry; Various methods and equipments; Arginase; Conductometric biosensor; Urease; l-Arginine determination; Quality-control analysis; Immobilization; Glutaral; Optimization; Arginine; Detection limit; Membrane; Ureas; Ungulata; Quantitative analysis; Bovine; Enzyme; Arginine determination; Sample; Serum albumin; Chemical sensor; Enzymatic method; Urea; Vertebrata; Sensitivity; Mammalia; Conductometry; Lysine; Storage stability; Biosensor; Quality control; Hydrolases; Relative error; Artiodactyla; Differential method; COMPLEX; UREASE; ELECTRODES; CHROMATOGRAPHY; L-Arginine determination; PLASMA; METABOLISM; AMINO-ACIDS; CAPILLARY-ELECTROPHORESIS; INHIBITORS; ARGINASE ACTIVITY
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2012.01.041

► Co-immobilization of arginase and urease allows a conductometric detection of l-arginine. ► A high sensitivity biosensor is based on cross-linking with glutaraldehyde. ► The biosensor is optimized for the real samples analysis. ► The biosensor remains to be functionally suitable over 3 months. A highly sensitive conductometric biosensor for l-arginine determination was developed by exploiting the unique biorecognition capacities of two enzymes of urea cycle – arginase (E.C. 3.5.3.1) and urease (E.C. 3.5.1.5). The enzymes were co-immobilized in a single bioselective membrane on the working sensor, while a lysine rich bovine serum albumin (BSA) membrane was immobilized on the reference sensor, allowing differential measurements. The optimum percentage ratio of arginase and urease within the bioselective membrane was determined when the biosensor sensitivity to l-arginine and urea was optimum. Analytical characteristics of the conductometric biosensor for l-arginine determination were compared for two types of enzyme immobilization (cross-linking with glutaraldehyde (GA) and entrapment in the polymeric membrane). The optimum features in terms of the sensitivity, the linear range, and the detection limit (4.2μS/mM, 0.01–4mM, and 5.0×10−7M, respectively) were found for l-arginine biosensor based on enzyme cross-linking with GA. A quantitative determination of l-arginine in the real sample (a drinkable solution “Arginine Veyron”) gave a satisfactory result compared to the data provided by the producer (a relative error was 4.6%). The developed biosensor showed high operational and storage stability.