Coupled reactions for the determination of analytes and enzymes based on the use of luminescence

✍ Scribed by Roda, A. ;Girotti, S. ;Ghini, S. ;Carrea, G.

• Publisher: John Wiley and Sons
• Year: 1989
• Weight: 763 KB
• Volume: 4
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170040157

✦ Synopsis

Immobilized enzymes are widely used i n the clinical laboratory in the assay of several analytes and enzymes. The use of immobilized enzymes makes these reagents recoverable and re-usable, and in most cases increases their stability and catalytic activity. In conjunction with bioluminescent enzymes (firefly and bacterial luciferases) and chemiluminescent catalyst (peroxidase) w e set up high-sensitive flow methods based on the use of nylon tube coil or epoxy methacrylate column as solid support.

All the NAD(P)/NAD(P)H-dependent dehydrogenases (bacterial luciferase), ATP-dependent enzymes (firefly luciferase) and oxidases producing HzOz (peroxidase) can be immobilized and a large variety of analytes have been sensitively measured. As an alternative format w e also reported a dry chemistry method in which all the enzymes, substrates and cofactors are ready to use, supported o n dry cellulose disks. Methodological problems such as flow conditions, stability, pH, ionic strength and analytical petformances are also reported.