RECOMBINANT HORSERADISH PEROXIDASE FOR ANALYTICAL APPLICATIONS

• Main Authors: А.M. Egorov, V. G. Grigorenko, I. P. Andreeva, M. YU. Ubtsova
• Format: Article
• Language: English
• Published: National Academy of Sciences of Ukraine and Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine. 2012-08-01
• Series: Biotechnologia Acta
• Subjects: recombinant horseradish peroxidase; fusion proteins; immunoassays; mediatorless bienzyme biosensor; Fab-fragments of antibody; heart-type fatty acid-binding protein; atrazine pesticides
• Online Access: http://biotechnology.kiev.ua/images/no4_2013/egorov_4_2013.pdf

The article deals with prospects of using recombinant horseradish peroxidase in analytical biochemistry and biotechnology. Problems of recombinant horseradish peroxidase cloning in different expression systems, possible approaches to their solution, advantages of recombinant recombinant horseradish peroxidase and recombinant horseradish peroxidase-fusion proteins for immunoassays are considered. Possibility for development of mediatorless bienzyme biosensor for peroxide and metabolites, yielding hydrogen peroxide during their transformations, based on co-adsorption of recombinant horseradish peroxidase and the appropriate oxidase was demonstrated. The possibility to produce a fully active recombinant conjugate of recombinant horseradish peroxidase with human heart-type fatty acid binding protein, which may be used in competitive immunoassay for clinical diagnosis of acute myocardial infarction, and recombinant conjugates (N- and C-terminus) of recombinant horseradish peroxidase with Fab-fragments of the antibody against atrazine, which may be applied for atrazine pesticides detection, are demonstra ted for the first time.