Tyrosinase Multilayer-Functionalised Carbon Nanotubes as Electrochemical Labels: Application To Immunoassay
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Publication Details
Author list: Chumyim P., Rijiravanich P., Somasundrum M., Surareungchai W.
Publisher: Springer
Publication year: 2014
Journal: BioNanoScience (2191-1630)
Volume number: 4
Issue number: 3
Start page: 240
End page: 250
Number of pages: 11
ISSN: 2191-1630
eISSN: 2191-1649
Languages: English-Great Britain (EN-GB)
Abstract
We have modified carboxylic acid-functionalised multi-walled carbon nanotubes (MWNTs) with multilayers of tyrosinase/polyallylamine using a layer-by-layer process. Based on optical determination, each tyrosinase deposition coated approximately three layers of enzyme onto the nanotubes. Based on kinetic analysis of the steady-state current of the modified MWNTs to catechol, the minimum value of k cat/K M for the immobilised enzyme was 3.9 ื 108 cm3 s-1 mol-1 ฑ 0.5 ื 108 cm3 s-1 mol-1 (n = 3), which is one order of magnitude less than the value for the enzyme in solution. As an illustration of an immunoassay application, the modified MWNTs were coated with streptavidin and then used as electrochemical labels for biotin-tagged anti-rabbit Salmonella polyclonal antibodies. Following optimisation of enzyme layer number, pH, incubation time and electrochemical parameters, heat-killed Salmonella Typhimurium cells were calibrated across the range 1 CFU ml-1 to 107 CFU ml-1 by direct enzyme-linked immunosorbent assay (ELISA) with a limit of detection (signal/noise = 3) of approx. 340 CFU ml-1. ฉ 2014 Springer Science+Business Media New York.
Keywords
Layer-by-layer, Tyrosinase
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