• Journal of Biosystems Engineering
• /
• v.28 no.2
• /
• pp.167-172
• /
• 2003

This study was performed to fabricate a batch-type SPR biosensing system using a thiolated E. coli antibody coupling, and to explore the feasibility of real-time detection of E. coii in a stagnant sample solution. In advance. “O” and “K” antigenic serotype E. coli antibodies were thiolated with sulfo-LC-SPDP and dithiothreitol, and immobilized by chemisorption in the gold surface of compact SPR sensors. When the SPR biosensor immobilized with E. coli antibody monitored a E. coli solution, it took 3 to 5 min to stabilize. The SPR biosensing system developed in this study was able to detect E. coli in the range above 10$^4$ CFU/mL at the 0.05 significant level. Also, the SPR biosensor had possibility to significantly detect E. coli in the range of 10$^2$ to 10$^4$ CFU/mL in E. coli solutions. Meanwhile, when the SPR biosensor immobilized with 5. coli antibody was cleaned with NaOH solutions, its ability to detect E. coli largely decreased due to wash-out of the immobilized antibody. In order to reuse the SPR sensor, it should be antibody-immobilized newly.

• Journal of Food Hygiene and Safety
• /
• v.18 no.3
• /
• pp.101-106
• /
• 2003

The aim of this study is to develop a label-free immunosensor for microbial detection and to evaluate its applicability to Pseudomonas aeruginosa detection in various food samples. The antibodies used were a polyclonal antiserum from rabbit (polyvalent type) and a monoclonal antibody raised against the flagella of P. aeruginosa. Antibody immobilization was done by a thiolated antibody chemisorption onto one gold electrode of a piezoelectric quartz crystal with a thiol-cleavable, heterobifunctional cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. To the Stomacher-treated samples from various raw and processed foods under cold storage, comprising sirloin, cod and pettitoes, spiking and enrichment culture were done to prepare the model samples, followed by the measurements of the frequency shifts after sample injections. The frequency shifts obtained by the sample matrices themselves were in the range of 52~89 Hz. The injections of the spiked samples caused the frequency shifts of 108~200 Hz, whereas the enriched samples decreased the steady-state resonant frequencies by 162~222 Hz. All sample measurements including baseline stabilization, sample injection and acquisition of the steady-state response were accomplished within 30 min.

• Applied Biological Chemistry
• /
• v.49 no.3
• /
• pp.254-258
• /
• 2006

Vitellogenin has been known as a potent biomarker protein for the estrogenic activity in fish exposed to endocrine disruptors. In this study, a piezoelectric immunosensor making use of an anticarp vitellogenin antibody and an AT-cut quartz crystal microbalance as the biological component and transducer was prepared, followed by its application to the analysis of carp vitellogenin as follows. Antibody immobilization was conducted by chemisorption of a thiolated antibody with a heterobifunctional thiolation cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. The reaction buffer for the immunosensor system was optimized as 0.1 M sodium phosphate (pH 7.4). Concentration-dependent sensor responses were obtained in the vitellogenin concentrations ranging from 0.4864 to 486.4000 nM, with a linear correlation between vitellogenin concentration and frequency shift in double-logarithmic scale. The limit of detection of the immunosensor for carp vitellogenin was presumed as 0.4864 nM.