Comparative study of controlled pore glass, silica gel and Poraver ® for the immobilization of urease to determine urea in a flow injection conductimetric biosensor system

• Published in: Biosensors & bioelectronics 2004-03, Vol.19 (8), p.813-821
• Main Authors: Limbut, Warakorn, Thavarungkul, Panote, Kanatharana, Proespichaya, Asawatreratanakul, Punnee, Limsakul, Chusak, Wongkittisuksa, Booncharoen
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Biosensor; Biosensors; Biotechnology; Coated Materials, Biocompatible - chemistry; Conductivity; Electrochemistry - instrumentation; Electrochemistry - methods; Enzyme Activation; Enzyme reactor; Enzymes, Immobilized - chemistry; Equipment Design; Equipment Failure Analysis; Flow Injection Analysis - instrumentation; Flow Injection Analysis - methods; Fundamental and applied biological sciences. Psychology; Glass - chemistry; Immobilized urease; Membranes, Artificial; Methods. Procedures. Technologies; Porosity; Protein Binding; Reproducibility of Results; Sensitivity and Specificity; Silica Gel; Silicon Dioxide - chemistry; Urea - analysis; Urea - chemistry; Urease - chemistry; Various methods and equipments
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2003.08.007

This study compared the responses of three enzyme reactors containing urease immobilized on three types of solid support, controlled pore glass (CPG), silica gel and Poraver ®. The evaluation of each enzyme reactor column was done in a flow injection conductimetric system. When urea in the sample solution passed though the enzyme reactor, urease catalysed the hydrolysis of urea into charged products. A lab-built conductivity meter was used to measure the increase in conductivity of the solution. The responses of the enzyme reactor column with urease immobilized on CPG and silica gel were similar and were much higher than that of Poraver ®. Both CPG and silica gel reactor columns gave the same limit of detection, 0.5 mM, and the response was still linear up to 150 mM. The analysis time was 4–5 min per sample. The enzyme reactor column with urease immobilized on CPG gave a slightly better sensitivity, 4% higher than the reactor with silica gel. The life time of the immobilized urease on CPG and silica gel were more than 310 h operation time (used intermittently over 7 months). Good agreement was obtained when urea concentrations of human serum samples determined by the flow injection conductimetric biosensor system was compared to the conventional methods (Fearon and Berthelot reactions). These were statistically shown using the regression line and Wilcoxon signed rank tests. The results showed that the reactor with urease immobilized on silica gel had the same efficiency as the reactor with urease immobilized on CPG.