• Journal of Sensor Science and Technology
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• v.20 no.6
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• pp.400-405
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• 2011

This study introduced the development of a strip type disposable enzyme-linked immunosensor platform for the detection of IgG. Strips of the strip sensor were fabricated by using commercial nitrocellulose filter membranes and a housing holder for the strips was manufactured by using a standard injection molding process for a plastic material. An IgG-urease conjugate was prepared and used for the competitive immune-binding with sample IgG. From the enzymatic reaction between the conjugated urease and urea added, ammonia was generated and caused a localized alkaline pH change on the immobilized antibody band which was coated onto the sensor strips. This pH increase subsequently caused a color change of the antibody band in the presence of a pH indicator, phenol red. Used in conjunction with a competitive immunoassay format, the intensity of the color produced is directly linked with the concentration of target analyte, IgG, and specific measurement of IgG in a lateral flow immunoassay format was achieved over the range 100 ppb to 2000 ppb IgG.

• KSBB Journal
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• v.24 no.3
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• pp.253-258
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• 2009

In this study, the optimization of fabrication conditions was accomplished to make immuno-strip biosensor by the combination of enzyme linked immunosorbent assay (ELISA) and immuno-chromatographic strip techniques for the detection of Escherichia coli O157 : H7. Optimal fabrication conditions of capture antibody concentration, detection antibody concentration, and additive composition of running buffer solution were determined. Optimal concentration was determined as 1.0 mg/mL for both of capture antibody and detection antibody. A composition of 0.5% Tween20 and 3% BSA were selected as optimal additive for buffer solution to prevent non-specific binding.

• Journal of Sensor Science and Technology
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• v.23 no.4
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• pp.278-283
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• 2014

In this study, we propose an immunosensor using zinc oxide nanorods (NRs) inside PDMS channel for detecting the influenza A virus subtype H7N9. ZnO with high isoelectric point (IEP, ~9.5) makes it suitable for immobilizing proteins with low IEP. In this proposed H7N9 immunosensor structure ZnO NRs were grown on the PDMS channel inner surface to immobilize H7N9 capture antibody. A sandwich enzyme-linked immunosorbent assay (ELISA) method with was used 3,3',5,5' tetramethylbenzidine (TMB) for detecting H7N9 influenza virus. The immunosensor was evaluated by amperometry at various H7N9 influenza antigen concentrations (1 pg/ml - 1 ng/ml). The redox peak voltage and current were measured by amperometry with ZnO NWs and without ZnO NWs inside PDMS channel. The measurement results of the H7N9 immunosensor showed that oxidation peak current of TMB at 0.25 V logarithmically increased from 2.3 to 3.8 uA as the H7N9 influenza antigen concentration changed from 1 pg/ml to 1 ng/ml. And then we demonstrated that ZnO NRs inside PDMS channel can improve the sensitivity of immunosensor to compare non-ZnO NRs inside PDMS channel.

• Journal of Microbiology and Biotechnology
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• v.13 no.4
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• pp.509-516
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• 2003

A model system for the immnunochemical detection of Escherichia coli O157:H7 using a combined immunomagnetic separation (IMS) and test-strip liposome immunoassay (LIA) procedure was developed. Immunomagnetic beads coated with anti-E. coli O157 IgG antibodies were used to separate the E. coli O157 (including the H7 serotype) from culture. Immunoliposomes, whose surface was conjugated to goat anti-E. coli O157:H7 IgG and which encapsulated the marker dye, sulforhodamine B, were used as a detection label. The test strip, onto which antibodies to goat IgG were immobilized, was the immunosensor capturing immunoliposomes that did not bind to E. coli O157:H7 on the immunomagnetic bead-E. coli O157:H7 complexes. In experiments, pure cell culture suspensions of $10^5 E.$ coli O157:H7 organisms per ml produced a measurable signal inhibition, whereas a weak yet detectable signal inhibition occurred with $10^3CFU/ml$. The inhibition signals increased, when the incubation time for IMS was extended to 90 min and higher IgG-tag density (0.4mol%) was used on the liposomes. With 0.2 and 0.4mol% IgG-tagged liposomes, the IMS-LIA procedure showed more improved signal inhibitions than those of a direct (no IMS) LIA. The combined assay, which measures the instantaneous signal from immunoliposomes, can be completed within 90 min, making it significantly faster than conventional plating methods and enzyme-linked immunosorbent assay (ELISA). Accordingly, it is quite feasible to use the combined immunoassay format of IMS and dye-loaded immunoliposomes for the detection of E. coli O157:H7.