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Title: Utilization of chip-based CE for avidin determination in transgenic tobacco and its comparison with square-wave voltammetry and standard gel electrophoresis

Author
item KRIZKOVA, SONA - MENDEL UNIV, BRNO, CZ
item HRDINOVA, VENDULA - MENDEL UNIV, BRNO, CZ
item ADAM, VOJTECH - MENDEL UNIV, BRNO, CZ
item BURGESS, ELISABETH - MT ALBERT RES CENTRE, NZ
item KRAMER, KARL - 5430-05-30 RETIRED
item MASARIK, MICHAL - MASARYK UNIV, BRNO, CZ
item KIZEK, RENE - MENDEL UNIV, BRNO, CZ
item Beeman, Richard

Submitted to: Chromatographia
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/18/2008
Publication Date: 4/18/2008
Citation: Krizkova, S., Hrdinova, V., Adam, V., Burgess, E., Kramer, K.J., Masarik, M., Kizek, R., Beeman, R.W. 2008. Utilization of chip-based CE for avidin determination in transgenic tobacco and its comparison with square-wave voltammetry and standard gel electrophoresis. Chromatographia. 67(S1): S75-S81. Doi: http://dx.doi.org/10.1365/s10337-008-0606-6.

Interpretive Summary: Analytical biosensors can be used to detect and quantify biologically important proteins in samples such as transgenic plants. Together with colleagues in the Czech Republic and New Zealand, we developed a microchip-based biosensor that measures the mobility of a charged protein in an electric field and utilized this sensor to measure levels of the biopesticide avidin in transgenic tobacco. This biosensor is more sensitive, rapid and reproducible than other analytical methods. It can be used by researchers to selectively monitor a range of biologically important compounds in different types of research and environmental samples.

Technical Abstract: Avidin transgenic plants are a potential tool for providing resistance against various species of insect pests due to the sequestration of vitamin H (biotin) in the plant from the insect pests. In this project we compared three techniques for avidin determination in transgenic tobacco plants, a novel chip-based capillary electrophoretic method (EXPERION), SDS-PAGE electrophoretic method and a square wave voltammetric method using a carbon past electrode. We determined that the automated capillary electrophoretic method is more rapid, sensitive, and reproducible than other methods. The avidin content measured in transgenic tobacco using chip-based capillary electrophoresis varied from 15 to 377 ng per mg depending on the individual plant, a result that was in good agreement with results obtained using the other methods.