Grafting odorant binding proteins on diamond bio-MEMS.
Odorant binding proteins (OBPs) are small soluble proteins found in olfactory systems that are capable of binding several types of odorant molecules. Cantilevers based on polycrystalline diamond surfaces are very promising as chemical transducers. Here two methods were investigated for chemically grafting porcine OBPs on polycrystalline diamond surfaces for biosensor development. The first approach resulted in random orientation of the immobilized proteins over the surface. The second approach based on complexing a histidine-tag located on the protein with nickel allowed control of the proteins' orientation. Evidence confirming protein grafting was obtained using electrochemical impedance spectroscopy, fluorescence imaging and X-ray photoelectron spectroscopy. The chemical sensing performances of these OBP modified transducers were assessed. The second grafting method led to typically 20% more sensitive sensors, as a result of better access of ligands to the proteins active sites and also perhaps a better yield of protein immobilization. This new grafting method appears to be highly promising for further investigation of the ligand binding properties of OBPs in general and for the development of arrays of non-specific biosensors for artificial olfaction applications.
CEA, LIST, Diamond Sensor Laboratory, 91191, Gif-sur-Yvette, France.
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R
Manai
ESIEE-Paris, ESYCOM University Paris-EST, Cité Descartes BP99, 93162 Noisy le Grand Cedex, France. Electronic address: emmanuel.scorsone@cea.fr.
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E
Scorsone
ESIEE-Paris, ESYCOM University Paris-EST, Cité Descartes BP99, 93162 Noisy le Grand Cedex, France.
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L
Rousseau
ESIEE-Paris, ESYCOM University Paris-EST, Cité Descartes BP99, 93162 Noisy le Grand Cedex, France.
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F
Ghassemi
ESIEE-Paris, ESYCOM University Paris-EST, Cité Descartes BP99, 93162 Noisy le Grand Cedex, France.
M
M
Possas Abreu
ESIEE-Paris, ESYCOM University Paris-EST, Cité Descartes BP99, 93162 Noisy le Grand Cedex, France.
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G
Lissorgues
The University of Manchester, School of Chemical Engineering and Analytical Science, Oxford Road, Manchester, M13 9PL, UK.
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N
Tremillon
The University of Manchester, School of Chemical Engineering and Analytical Science, Oxford Road, Manchester, M13 9PL, UK.
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H
Ginisty
CEA, LIST, Diamond Sensor Laboratory, 91191, Gif-sur-Yvette, France.
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JC
Arnault
The University of Manchester, School of Chemical Engineering and Analytical Science, Oxford Road, Manchester, M13 9PL, UK.
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E
Tuccori
GTP Technology, Immeuble Biostep, 436 Rue Pierre et Marie Curie, 31670 Labège, France.
M
M
Bernabei
GTP Technology, Immeuble Biostep, 436 Rue Pierre et Marie Curie, 31670 Labège, France.
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K
Cali
GTP Technology, Immeuble Biostep, 436 Rue Pierre et Marie Curie, 31670 Labège, France.
K C
KC
Persaud
CEA, LIST, Diamond Sensor Laboratory, 91191, Gif-sur-Yvette, France.
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P
Bergonzo
D040761:Biomimetic Materials; D018130:Diamond / Q000737:chemistry*; D058266:Dielectric Spectroscopy / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D055617:Micro-Electrical-Mechanical Systems / Q000295:instrumentation*; D009812:Odorants / Q000032:analysis*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000295:instrumentation; D018035:Receptors, Odorant / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012903:Smell
D018035:Receptors, Odorant; C045648:odorant-binding protein; D018130:Diamond
Artificial olfaction; Diamond cantilevers; Odorant binding protein