title: Construction of a peptide with an electroactive daunomycin like a pendant arm to detect ovalbumin.
abstract: In this study, a peptide-1 (RNRCKGTDVQAW) constructing lysozyme was conjugated with an electroactive daunomycin in order to voltammetrically detect ovalbumin (OVA). Hetero-bifunctional cross-linking agents with four kinds of ethylene chains in differing lengths were used to bind the peptide-1 and daunomycin. After a cross-linking agent had reacted with an amino group of daunomycin, the compound was introduced into the peptide to the cysteine residue in the peptide using a pendant arm. The OVA was sensed via a change in the electrode response of the daunomycin moiety, based on the binding between the peptide and the OVA. The adsorption of the peptide probe on the electrode increased with increases in the ethylene chain. The binding constants between the peptide probes and the OVA, however, did not depend on the length of the chain. This was because the ethylene chain influenced the binding. When the peptide and the daunomycin were bound using N-(6-maleimidocaproyloxy) sulfosuccinimide, the electrode response of the peptide probe was the most sensitive from among the four cross-linking agents. The calibration curve of the OVA using the peptide probe was linear and ranged from 1.5×10(-11) to 3.0×10(-10)M. Furthermore, this method could be applied to the electrochemical sensing of the OVA in egg whites and in fetal bovine serum.
authors.affiliation: Maebashi Institute of Technology, Gunma 371-0816, Japan. Electronic address: kzsuga@maebashi-it.ac.jp.
authors.forename: Kazuharu
authors.initials: K
authors.lastname: Sugawara
authors.affiliation: Maebashi Institute of Technology, Gunma 371-0816, Japan.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Kadoya
authors.affiliation: Department of Environmental Biology and Chemistry, Graduate School of Science and Engineering for Research, University of Toyama, Toyama 930-8555, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Kuramitz
mesh_terms: D000818:Animals; D002417:Cattle; D002642:Chick Embryo; D002645:Chickens; D003432:Cross-Linking Reagents / Q000737:chemistry; D003630:Daunorubicin / Q000737:chemistry*; D004529:Egg White / Q000032:analysis / Q000737:chemistry; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D013058:Mass Spectrometry; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D010047:Ovalbumin / Q000032:analysis* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D011784:Quail; D012680:Sensitivity and Specificity; D044967:Serum / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D015335:Molecular Probes; D010455:Peptides; D010047:Ovalbumin; D009113:Muramidase; D003630:Daunorubicin
keywords: Alkyl chain; Binding constant; Ovalbumin; Peptide probe