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Paper
Analyst, 2009, 134, 926 - 932, DOI: 10.1039/b813451k
High performance immunoassay using immobilized enzyme in nanoporous carbon
Yunxian Piao, Dohoon Lee, Jungbae Kim, Jaeyun Kim, Taeghwan Hyeon and Hak-Sung Kim
A highly stable immunoassay format was constructed using signal-generating enzyme immobilized in nanoporous carbon. A mesocellular carbon foam, called MSU-F-C, was loaded with horseradish peroxidase (HRP), followed by cross-linking of the enzyme using glutaraldehyde (GA) and modification of the surface with anti-human IgG through EDC/sulfo-NHS chemistry. The resulting MSU-F-C/HRP/anti-human IgG stably retained immobilized enzymes and antibodies, showing higher thermal stability. The MSU-F-C/HRP/anti-human IgG retained about 80 % of initial enzyme activity at 40 °C after a 5 h incubation, while the HRP/anti-human IgG conjugate resulted in almost 90% loss of initial activity in the same condition. In bead-based immunoassays, the signal amplification using MSU-F-C/HRP/anti-human IgG enabled the sensitive colorimetric detection of a target analyte, human IgG, in a detection limit of
33 pM, with negligible cross-reactivity against rabbit and chicken IgGs.
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