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Title: APPLICATION OF AVIDIN-BIOTIN TECHNOLOGY AND ADSORPTIVE TRANSFER STRIPPING SQUARE-WAVE VOLTAMMETRY FOR DETECTION OF DNA HYBRIDIZATION AND AVIDIN IN TRANSGENIC AVIDIN MAIZE

Author
item MASARIK, MICHAL - ACAD SCI CZECH REPUBLIC
item KIZEK, RENE - ACAD SCI CZECH REPUBLIC
item Kramer, Karl
item BILLOVA, SABINA - ACAD SCI CZECH REPUBLIC
item BRAZDOVA, MARIE - ACAD SCI CZECH REPUBLIC
item VACEK, JAN - ACAD SCI CZECH REPUBLIC
item BAILEY, MICHELE - PRODIGENE, INC
item JELEN, FRANTISEK - ACAD SCI CZECH REPUBLIC
item HOWARD, JOHN - PRODIGENE, INC

Submitted to: Analytical Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/13/2003
Publication Date: 6/1/2003
Citation: MASARIK,M., KIZEK,R., KRAMER,K.J., BILLOVA,S., BRAZDOVA,M., VACEK,J., BAILEY,M., JELEN,F., HOWARD,J.A., APPLICATION OF AVIDIN-BIOTIN TECHNOLOGY AND ADSORPTIVE TRANSFER STRIPPING SQUARE-WAVE VOLTAMMETRY FOR DETECTION OF DNA HYBRIDIZATION AND AVIDIN IN TRANSGENIC AVIDIN MAIZE, ANALYTICAL CHEMISTRY 75: 2663-2669. 2003.

Interpretive Summary: Because of new developments in biotechnology, novel highly sensitive analytical methods are needed to detect or monitor DNA and proteins produced by genetically modified organisms in both the laboratory and in the field. In collaboration with scientists at the Academy of Sciences of the Czech Republic and Prodigene, Inc., an agricultural biotechnology company, we describe here an electrochemical method based on avidin-biotin biotechnology that is very sensitive and specific for nucleic acids and recombinant proteins produced by transgenic crops. This method might be useful for monitoring fields surrounding transgenic crops for unintentional contamination of other crops and weedy species.

Technical Abstract: The proteins streptavidin and avidin were electrochemically detected in solution by adsorptive transfer stripping square wave voltammetry (AdTS SWV) at a carbon paste electrode (CPE). AdTS SWV was used to quantify biotinylated oligonucleotides, DNA hybridizations, and avidin in extracts of transgenic avidin maize. The detection limits of denatured and native streptavidin were 6 pM and 120 nM, respectively. The results demonstrated that streptavidin/avidin AdTS SWV is a sensitive and specific method for quantifying DNA and proteins in biological samples such as foods and tissue extracts, including genetically modified crops (avidin maize) and other plants in neighboring fields.