NDLI: Peptide conjugated chitosan foam as a novel approach for capture-purification and rapid detection of hapten--example of ochratoxin A.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Hassine, A. Ibn Had Gonzalez, C. Bazin, I. Soleri, R. Demey, H. Guiseppi-Elie, A. Tria, S. A.
Description	Country affiliation: France Author Affiliation: Soleri R ( Ecole des mines d'Alès, LGEI, 6 avenue de Clavière, 30319 Alès Cedex, France.); Demey H ( Ecole des mines d'Alès, LGEI, 6 avenue de Clavière, 30319 Alès Cedex, France.); Tria SA ( Ecole des mines d'Alès, LGEI, 6 avenue de Clavière, 30319 Alès Cedex, France.); Guiseppi-Elie A ( Clemson University, C3B, 100 Technology Drive, Clemson, SC 29634, USA.); Hassine AI ( Ecole des mines d'Alès, LGEI, 6 avenue de Clavière, 30319 Alès Cedex, France.); Gonzalez C ( Ecole des mines d'Alès, LGEI, 6 avenue de Clavière, 30319 Alès Cedex, France.); Bazin I ( Ecole des mines d'Alès, LGEI, 6 avenue de Clavière, 30319 Alès Cedex, France. Electronic address: ingrid.bazin@mines-ales.fr.)
Abstract	A novel bioassay for the detection and monitoring of Ochratoxin A (OTA), a natural carcinogenic mycotoxin produced by Aspergillus and Penicillium fungi, has been developed and applied for the screening of red wine. Here we report the immobilization and orientation of NOF4, a synthetic peptide, onto 3-D porous chitosan supports using a N-terminal histidine tag to allow binding to M(++) ions that were previously adsorbed onto the high surface area biopolymer. Three divalent cations (M(++)=Zn(++), Co(++), Ni(++)) were evaluated and were found to adsorb via a Langmuir model and to have binding capacities in the order Zn(++)>Co(++)>Ni(++). Following Zn(++) saturation and washing, C-terminus vs. the N-terminus His-tagged NOF4 was evaluated. At 1000 µg L(-1) OTA the N-terminus immobilization was more efficient (2.5 times) in the capture of OTA. HRP labeled OTA was added to the antigen solutions (standards or samples) and together competitively incubated on biospecific chitosan foam. The chemiluminescence substrate luminol was then added and after 5 min of enzymatic reaction, light emission signals (λmax=425 nm) were analyzed. Calibration curves of %B/B0 vs. OTA concentration in PBS showed that half-inhibition occurred at 1.17 µg L(-1), allowing a range of discrimination of 0.25 and 25 µg L(-1). In red wine, the minimum concentration of OTA that the system can detect was 0.5 µg L(-1) and could detect up to 5 µg L(-1). Assay validation was performed against immunoaffinity column (IAC) tandem reversed-phase high pressure liquid chromatography with fluorescence detection (HPLC-FLD) and provided quite good agreement. The association of chitosan foam and specific peptide represents a new approach with potential for both purification-concentration and detection of small molecules. In the future this assay will be implemented in a solid-sate bioelectronic format.
ISSN	09565663
Volume Number	67
e-ISSN	18734235
Journal	Biosensors and Bioelectronics
Language	English
Publisher	Elsevier
Publisher Date	2015-05-15
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Biosensing Techniques Haptens Isolation & Purification Ochratoxins Chitosan Chemistry Chromatography, High Pressure Liquid Histidine Peptides Journal Article Research Support, Non-u.s. Gov't Discipline Biotechnology
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry

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