Development of a chemiluminescent immunosensor for chloramphenicol

• Published in: Analytica chimica acta 2006-09, Vol.578 (1), p.19-24
• Main Authors: Park, In-Seon, Kim, Namsoo
• Format: Article
• Language: English
• Subjects: Chloramphenicol detection; Direct competitive chemiluminescent immunosensor; Model sample analysis; System optimization
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2006.07.015

A direct competitive chemiluminescent immunosensor system that exploits the competition between chloramphenicol (CAP) as an analyte and CAP–horseradish peroxidase conjugate as a tracer for binding to an anti-CAP antibody on a solid support was devised by installing a flow-through cell which was connected to an injector and a peristaltic pump inside a dark box, followed by positioning a photomultiplier tube as light detector in front of it. The anti-CAP antibody was immobilized onto positively charged Biodyne B membrane pieces by a dipping procedure. The operating conditions for the immunosensor were selected with respect to substrate composition (0.25, 13.3 and 0.66 mM for luminol, H 2O 2 and p-iodophenol, respectively), injection volume of the substrate solution (200 μL) and the concentrations of antibody for immobilization (0.10 mg mL −1) and tracer (0.030 mg mL −1). At these conditions, sensor response according to analyte concentration was well fitted to a linear equation when plotted in semi-logarithmic scale, with the limit of detection for CAP of 10 −8 M. By using the immunosensor, CAP measurement in the model samples prepared from five food materials was conducted.