title: Use of recombinant oestrogen binding protein for the electrochemical detection of oestrogen.
abstract: We have previously reported the development of an electrochemical method to quantitatively detect vertebrate oestrogens using an oestrogen binding protein (EBP1) present in wild type Saccharomyces cerevisiae and Candida albicans cells. However, the assays were complex and slow with both whole cells and cell lysate. In this work we report the transfer of the EBP1 gene to an industrial yeast, the addition of a his tag sequence to simplify purification of the protein, and the oestrogen binding characteristics of the protein. The recombinant protein (Ebp1p-6h) can now be produced by a non-pathogenic cell, and has shown good stability both when in use and when lyophilised. The detection range covers likely environmental concentrations of free oestrogens and the limit of detection is below the environmental concentration that has significant biological effect. In addition the assay period has been reduced to approximately 2min. This work reports progress toward the construction of a rapid, portable oestrogen sensor that is not restricted to use to the laboratory.
authors.affiliation: School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand.
authors.forename: Vimal
authors.initials: V
authors.lastname: Vijayan
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Giersberg
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Chamas
authors.affiliation: School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand.
authors.forename: Mamta
authors.initials: M
authors.lastname: Mehrotra
authors.affiliation: Department of Biosciences and Bioengineering, Indian Institute of Technology (Bombay), Mumbai, India.
authors.forename: Venkata
authors.initials: V
authors.lastname: Chelikani
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany.
authors.forename: Gotthard
authors.initials: G
authors.lastname: Kunze
authors.affiliation: School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand. Electronic address: keith.baronian@canterbury.ac.nz.
authors.forename: Keith
authors.initials: K
authors.lastname: Baronian
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002176:Candida albicans / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D004958:Estradiol / Q000032:analysis* / Q000378:metabolism; D004967:Estrogens / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D011960:Receptors, Estrogen / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D004967:Estrogens; D011960:Receptors, Estrogen; D011994:Recombinant Proteins; C010140:estrophilin; D004958:Estradiol
keywords: Electrochemical detection of 17β-oestradiol binding; Recombinant yeast oestrogen binding protein; SPR detection of 17β-oestradiol binding