Stable Immobilization of Anti-Beta Casein Antibody onto Layered Double Hydroxides Materials for...

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REVIEW

Copyright copy 2009 American Scientific PublishersAll rights reservedPrinted in the United States of America

SENSOR LETTERSVol 7 647ndash655 2009

Stable Immobilization of Anti-Beta Casein Antibodyonto Layered Double Hydroxides Materials

for Biosensor Applications

Z M Baccar1lowast S Hidouri1 N El Bari2 N Jaffrezic-Renault3 A Errachid3 and N Zine41UR-PCGM National Institute of Research in Physics and Chemistry Analysis (INRAP) 2020 Sidi Thabet Tunisia

2U R Biotechnology Food Agronomy and Biomedical Analysis Faculty of Science MeknesMoulay Ismail University PO Box 11201 Zitoune 50014 Meknes Morroco

3University of Lyon Laboratory of Analytical Sciences UMR CNRS 5180 University of Claude Bernard Lyon I France4Laboratori de Nanobioenginyeria Institut de Bioenginyeria de Catalunya (IBEC) Baldiri Reixac 10-12 08028 Barcelona Spain

(Received 15 January 2009 Accepted 31 July 2009)

This review presents the development of new kind of antibodyLDH (layered double hydroxides)hybrid nanomaterials for beta casein detection The preparation method of the LDH is described It isbased on the co-precipitation of metallic salts in constant pH and temperature The chosen LDH arehydrotalcites (Mg2AlCO3 Mg3AlCO3) Zaccagnaite Zn2AlCO3 and hydrocalumite Ca2AlCl Finallythe antibody is immobilized into the LDH materials using Layer-by-Layer method by autoassemblyIn this work we studied the surface properties of the prepared hybrid biomembranes using X-raydiffraction Infrared spectroscopy in ATR mode and Atomic Force Microscopy (AFM) These tech-niques allow describing the antibody immobilization and its interactions with LDH The antibodywas adsorbed and its morphology was conserved in its new environment after more than 15 dayscontinuously in PBS solution promising a constant biosensor performance

Keywords Anti -Casein Antibody Antibody Immobilization LDH Hybrid Biomaterials UreaBiosensors

CONTENTS

1 Introduction 6472 Experimental Procedures 649

21 Preparation of Antibody and Antigen 64922 Preparation of LDHs 65023 Fabrication of InsulatorSemiconductor (IS) 65024 Anti CaseinLDH Hybrid Material Deposition 65025 Surface Characterizations 650

3 Results and Discussion 65031 XRD Characterization of LDH Powders 65032 Surface Characterizations of the Hybrid

CaseinLDH 6514 Conclusion 654

Acknowledgments 654References and Notes 654

1 INTRODUCTION

During the past two decades numerous serious sanitaryfood problems appeared in developed country Because of

lowastCorresponding author E-mail Zouhair_baccartopnettn

the government restriction in industrial activities and espe-cially the food industry the monitoring and the continuouscontrol of process the analysis of production are requiredThe developed systems of monitoring based on real timesensor are needed For this reason an extensive researchhas been carried out to improve new materials for biolog-ical species immobilization In fact the specificity and theperformance of the biosensor depend on the bioreceptorand its conformation into host material on the tranducerAn essential requisite for the development of biosensor isthe immobilization of biomolecule by a method whereinthe bioreceptor remains active1 The standard biomoleculeimmobilization are based on physical adsorption covalentbinding cross-linking and entrapment into polymers orsolgel matrices2ndash9 In fact during the process of immobi-lization or intercalation solvent used for opening the hostmaterials can denature or raceme the bioreceptor causingits reduction or lost of activity10 In order to reduce thisrisk new materials have been studied for the biologicalentrapment and adsorption A new generation of func-

tionalized materials is biomoleculeLDH hybrid materials

Sensor Lett 2009 Vol 7 No 5 1546-198X20097647009 doi101166sl20091129 647


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Stable Immobilization of Anti-Beta Casein Antibody onto LDH Materials for Biosensor Applications Baccar et al

Z M Baccar received his PhD in Electronic devices from Ecole Centrale de Lyon Francein 1996 He was assistant professor in University of Monastir Tunisia in 1996 than inuniversity of 7 Nov in Carthage in 2000 Since 2008 he joined the National Institute ofResearch in Physics and Chemistry Analysis in Sidi Thabet as a researcher His researchactivities consist on the development and the preparation of membranes for biosensors(ISFETs ISEs ) based on hybrid nanomaterials He is leader in 4 Spanish-Tunisian(AECI and AECID) and one Morocco-Tunisian (CMPTM) program on development of sen-sors for biomedical and environmental applications

S Hidouri received his bachelor degree from University of 7 Nov of Carthage the facultyof Sciences of Bizerte (FSB) in 2004 He received the Theoretical master in Biocomputerfrom the High School of Computer Science (ENSI) in 2005 In 2009 he received a secondmaster in hydrobiology from the University of 7 Nov of Carthage the faculty of Sciencesof Bizerte (FSB) Actually he is a PhD student

N El Bari was awarded a PhD in Biology from the University Moulay Ismail (MeknesMorocco) in 1995 Since 1999 she has been a professor at the Faculty of Sciences in Meek-ness and is currently head of the Biotechnology Agroalimentary and Biomedical AnalysisGroup One of her research interest is the application of electronic nose techniques to foodspoilage

N Jaffrezic-Renault received her engineering degree from the Ecole Nationale Supeacuterieurede Chimie Paris in 1971 and the Doctorat drsquoEtat eacutes Sciences Physiques from the Uni-versity of Paris in 1976 She joined Ecole Centrale de Lyon France in 1984 and ClaudeBernard University Lyon 1 in 2007 As Director of Research at the Centre National de laRecherche Scientifique and the president of the chemical micro sensor club (CMC2) herresearch activities include the preparation and physicochemical characterization of mem-branes for chemical sensors (ISFETs optical fiber sensors electrochemical sensors) She iscurrently leading 6 PhD students 2 MSc These students are involved in one Europeanproject on microsystem for diagnostics and in several national projects for the developmentof microsystems for environmental monitoring and for food safety She is leader in a nationalprogram (INTEGREAU) involving 4 academic partners and 2 companies for the develop-

ment and industrial transfer of a generic microsystem for the monitoring of natural waters according to the frameworkdirective on water quality She published more than 350 papers with around 4000 citations

648 Sensor Letters 7 647ndash655 2009


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REVIEW

Baccar et al Stable Immobilization of Anti-Beta Casein Antibody onto LDH Materials for Biosensor Applications

A Errachid was born in Khenifra Morocco in 1966 He graduated in Physics from theUniversity M Ismail Meknes 1992 and received a PhD degree in Electronic Engineering(with honors) from the Universitat Autogravenoma de Barcelona Spain in 1998 From 1998to 2001 he worked at the Centro Nacional de Microelectroacutenica (CNM) as researcher Atthe end of 2001 he joined the Research Centre for BioElectronics and NanoBioScience(CBEN) at the University of Barcelona as researcher within the Spanish Ramon y Cajal Pro-gram He has been working in the design fabrication and characterisation of chemical- andbio-sensors based on microelectrodes and FET (Field-Effect-Transistor) and in the develop-ment of integrated instrumentation for ISFETs (Ion-sensitive Field-Effect-Transistor) He hasbeen involved in several European Projects (DEMAC BARMINT MICRO-CARD MICRO-TRANS SPOT-NOSED MAPTECH DVT-IMP) From 2006 to 2008 he was working as a

senior researcher at the Institute for Bioengineering of Catalonia (IBEC) at the University of Barcelona At the end of2007 he joined Claude Bernard University-Lyon 1 as a Professor His current research activity is focussed in BioElec-tronics Biofonctionalisation and NanoBiotechnology

N Zine was born in Meknes Morocco in 1969 She graduated in Chemistry from the Uni-versity M Ismail Meknes in 1994 She received a Master in Material Science in 1997 anda PhD degree in Chemistry in 2005 both from the ldquoUniversitat Autogravenoma de BarcelonardquoSpain She worked at Barcelona Microelectronic Institute (IMB-CSIC) as a post-doctoralresearcher From 2006 she is working in the Barcelona Scientific Park within the Span-ish ldquoJuan de la Ciervardquo Program Shersquos author and co-author of 16 papers and more than28 communications in scientific meetings Her research interest is the Chemical Charac-terization of Silicon Chemical Sensors and Biosensors Specially of ISFET devices andmicroelectrodes

Layered double hydroxide (LDH) are anionic clays definedby a typical formula structure [M2+

1minusX M3+X (OH)2]

x+ middot[Axq]

qminus middot nH2O1112 where M2+ and M3+ are di- and

trivalent cations including Mg2+ Fe2+ Ni2+ Cu2+ Co2+Mn2+ Zn2+ or Cd2+ and Al3+ Cr3+ Ga3+ or Fe3+ respec-tively and A is an anion In the LDH isomorphousreplacement of a fraction of the divalent ions with a triva-lent cation such as Al3+ induces a positive charge on thelayers [M2+

1minusX M3+X (OH)2] which necessitates the pres-

ence of interlayer balancing anion charge for a result-ing neutral charge of the LDH materials This class ofinorganic materials can be considered as a suitable plat-form for biomolecular immobilization13ndash15 for biosensorsapplication16ndash18 bioreactor19 drugs formulation or medici-nal therapy20ndash26 It presents a lot of advantages such as thebi-dimensional structure and its high opening and anioniccapacity of exchange27 its high ability to encapsulateadsorb or intercalate biomolecules28ndash30 into positive chargelayers orand interfoliar layer3132 its biocompatibility Finally these inorganic materials are prepared in soft con-dition with a soft chemical process15 and the risk of denat-uration of the biomolecule is not significant and can beconsideredIn this paper we attempt to prepare antibodyLDHs

materials for beta casein biosensor application We pre-pared and we studied the adsorption of anticasein betathrough five prepared LDHs hydrotalcites (Mg2AlCO3Mg3AlCO3) hydrocalumite (Ca2AlCl) and Zaccagnaite

(Zn2AlCO3) The LDHs and the hybrid antibodyLDHswere characterized by X-ray Diffraction IRTF-ATR spec-troscopy and AFM imaging We followed the evolutionand the antibody linked materials after 15 daysrsquo stay inanalytical solutions This approach allows proving the fea-sibility of performing a beta casein-biosensor using a softprotocol of probe immobilization with a good conforma-tion and performance

2 EXPERIMENTAL PROCEDURES

21 Preparation of Antibody and Antigen

211 Purification of -Casein

-casein purification were performed using two comple-mentary processes preliminary preparation of prepurified-casein in high quantity based on Aschaffenburg proto-col (1963) 33 modified by Garnier and al (1964) 34 followedby chromatographic method by ion exchange for enhancethe purified product

212 Quantification and Control of Preparation

Finally the monitoring and the control of the both -casein(prepurified and purified) were performed by Polyacry-lamide gels electrophoresis according to the procedureadapted for PhastSystem35 The obtained electrophorgram

Sensor Letters 7 647ndash655 2009 649


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EVIEW


shows a well purified product (715 of prepurified-casein and 899 of purified -casein)

213 Preparation of Antibody

Anti--casein antiserum was obtained by immunizing rab-bits with intradermal injection of purified -casein (10 mgin 1 mL of 014 M NaCl) emulsified with 1 mL of Freundrsquoscomplete Adjuvant (Behring Marburg FRG) for the firstinjection and incomplete adjuvant for following boosterinjection (after 13 36 65 90 days) antisera were collectedafter 7 days after each booster injection Reactivity andspecificity of antisera were determined by the method ofOuchterlony (Immunodiffusion double)

22 Preparation of LDHs

Three types of LDH (hydrotalcite hydrocalumite andzaccagnaite) were prepared by co-precipitation at con-stant pH and temperature previously described36 For allpreparation the protocol is similar only reagent pHand temperature condition are different For hydrotalcite(respectively Zaccagnaite) synthesis we added drop todrop 01 M solutions of AlCl3 and MgCl2 (resp ZnCl2) atroom temperature The pH of the mixture was maintainedat pH 80 by adding a basic mixture solution of NaOH2 M and NaCO3 0125 M for hydrotalcite (resp 05 M forZaccagnaite) The final solution were washed with MilliQwater filtered then dried for 12 hrs at 100 C and finallycrushed For hydrocalumite the protocol is similar how-ever the divalent metallic salt is replaced by CaCl2 theexperimental conditions for synthesis are 65 C and pHwas maintained at 115 by adding 2 M NaOH solution aspreviously described37

23 Fabrication of InsulatorSemiconductor (IS)

The microelectronic manufacturing process for planardevices was performed at Centro National de Microelec-troacutenicagrave (CNM) The substrate sample used in this workis 1 cmtimes 1 cm of Si3N4SiO2Si(p-type) structure TheirInsulatorSemiconductor (IS) samples are performed in100 mm diameter wafers of 100-oriented p-SiO2 withresistivity of 4ndash40 middotcm The process started with a ther-mal oxidation process to grow a silicon dioxide layer (SiO2

78 nm) on silicon wafers in a hydrox furnace at 950 CThen an LPCVD Si3N4 layer (100 nm) was deposited onsilicon dioxide surface at 800 C

24 Anti CaseinLDH Hybrid Material Deposition

Hybrid materials were prepared by Layer-by-Layer auto-assembling LDH and anti casein layers onto the IS struc-ture The surface of sample was firstly cleaned usingPIRANHA mixture After that a drop of approximately10 l of dispersed LDH in MilliQ water (25 mgml) was

deposited on the IS surface and spinned during 40 s attwo different speeds (1000 trmin during the first 10 s thenat 4000 trmin during the last 30 s) After that a dropof approximately 10 l of concentrated anti-casein wasdeposited on surface and left at 4 C in a saturated PBSatmosphere over the night Finally the non-bonded anti-bodies were removed by washing samples with phosphatebuffer at pH 74 After experiment the samples werestored in PBS solution at 4 C

25 Surface Characterizations

Powder X-ray Diffraction (XRD) patterns were performedon a Panalytical XrsquoPert Pro diffractometer using Co K1(40 kV 30 mA) radiation and continued scanning modeThe patterns were recorded in a 2 range from 7 to 80in steps of 016 and counting time of 2 s per stepAbsorbing Infrared Fourier Transformed (IRTF) spec-

tra in ATR mode were recorded using an EQUINOX55 (Bruker) spectrophotometer in the range of 4000ndash550 cmminus1 with 2 cmminus1 resolution and averaging 128 scansAFM Imaging was performed in tapping mode using a

Dimension 3100 (Veeco) Atomic Force Microscope

3 RESULTS AND DISCUSSION

31 XRD Characterization of LDH Powders

As shown in Figure 1 the XRD patterns confirm the crys-talline phase identity of the synthesized LDH36 The ZnxAland Ca2AlCl XRD patterns present symmetrical peaksAs resumed in Table I obtained experimental crystallo-graphic properties of prepared Ca2AlCl correlate with the-oretical value given in ICSD file For MgxAl patternsasymmetrical peaks were obtained and were in correlationwith the well-crystallized CO3 hydrotalcite structure38

0 20 40 60 80

(-12

7)

(006

)

(040

)

(-12

3)

(003

)

2θ (deg)

Ca2Al

Rel

ativ

e in

tens

ity (

)

Zn2Al(116

)(0

113)

(113

)(1

10)

(011

1)

(101

0)

(018

)(1

07)(0

15)

(012

)(1

01)

(006

)

(018

)

(015

)

(012

)

(110

)(1

13)(003

)

Mg2Al

(202

6)(2

23)

(111

8)

(021

0)

(001

8)

(001

2)

(006

)

Mg3Al

(002

)

(-44

6)(2

46)

(206

)(044

)(3

23)

(400

)

(004

)

Fig 1 XRD diffractograms of LDH powder



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REVIEW


Table I Indexing XRD patterns for the prepared Ca2AlCl LDH powderand comparison to the model compounds retried in ICSD data base

Ca2Al

Space group MonoclinicChemical composition Ca2Al(OH)6Cl 2H2Oa (nm) 1002b (nm) 1151c (nm) 1286FWHM (2) 00929

dspacing (nm)

Most intensive peak list (h k l) dexperimental (nm) dModelling (nm)1st Peak (0 0 2) 0781 07902nd peak (0 0 4) 0391 03953rd peak (minus1 2 3) 0379 0385

32 Surface Characterizations of theHybrid CaseinLDH

We studied the affinity of the positively charged layersof LDH for the anticasein and their capability as linkedmaterial for the antibodies This study was carried out bycoupling IR-spectroscopy in ATR mode and AFM mor-phology scanning in tapping mode

321 IR Characterization in ATR Mode

The result obtained by ATR-Infra red spectroscopyexhibit vibration of antibodyLDH materials As shownin Figure 2 all IR spectra of antibodyLDHs are simi-lar in shape This is probably due to the fact that anti-bodyLDH interaction depends on hydroxyls groups ofpositively charged layers The observed large band at2700ndash3150 cmminus1 was assigned to intermolecular chelateH binding Another band is obtained at 2560ndash2650 cmminus1It can be due to stretching OH vibrations The observed1974ndash2029 cmminus1 band correspond to amid bands Whilethe peaks obtained at 2029 cmminus1 can be associated to thesecondary amide band CndashNO2 stretching vibrationFinally the spectra obtained with anti caseinLDHs

show a band at 1140 cmminus1 and a peak at 980 cmminus1 they canbe assigned to an anhydride CndashOndashC stretching vibrationrespectively to a secondary aliphatic amine CndashN stretchingvibrationAll Infra red patterns of antibody-LDH shows peaks

at 2367ndash2346 cmminus1 They can be assigned to anhydrideC C stretching vibration However the antibodyMgxAlspectra show peak at 2162 cmminus1 that are assigned to C Ostretching vibration40 The corresponding symmetry andasymmetric CndashOndashC stretching vibration are observed at1300 resp 1150 cmminus1 The obtained peak at 668ndash618 cmminus1

is due to NndashH stretching vibrationIn conclusion the immobilization of the antibody is due

to the interaction between the polar functions and the OHgroup of the LDHs in the first time and amine function and

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

ν (cmndash1)

(a)Mg2AlMg2Alab1jMg2Alab10j

ν (cmndash1)

(b)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

Ant

i sym

DO

Mg3AlMg3Alab1j

Mg3Alab10j

Carboxylate 0CO-

sym

2700-3150

Intermolecular chelate hdrogen bonds

2650-2560

streatching thiols SH

ν (cmndash1)

(c)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

CaAl

Ca2alab10jCa2alab1j

ν (cmndash1)

(d)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

Zn2AlZn2Alabj1Zn2Alabj10

Fig 2 IR spectra of LDH and their anti-caseinLDH evolution dur-ing the time of analyst exposure (a) anti-caseinMg2Al (b) -anticaseinMg3Al (c) anti-caseinCa2Al d) anti-caseinZn2Al



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EVIEW


Initial β anti-caseinCa2Al morphology

(a)

Initial β anti-caseinZn2Al morphology

Initial β anti-caseinMg2Al morphology Initial β anti-caseinMg3Al morphology

(b)

β anti-caseinCa2Al 72 h in PBS β anti-caseinMg2Al 72 h in PBS

Fig 3 Continued



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REVIEW


β anti-caseinMg3Al 72 h in PBS β anti-caseinZn2Al 72 h in PBS

β anti-caseinZn2Al 170 h in PBS

(c)

β anti-caseinMg3Al 170 h in PBS

β anti-caseinMg2Al 170 h in PBS β anti-caseinCa2Al 170 h in PBS

Fig 3 AFM images of the surface topography of the hybrid anti caseinLDH (Surface scan 1 mtimes 1 m) The follow-up of the morphologyafter storage in physiological PBS solution at room temperature (a) LDH (b) initial anti bodyLDH (b) after 72 hours in PBS buffer (c) after 170 hin PBS buffer



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EVIEW


Table II The resumed roughness value of the different hybrid biomem-branes and their follow up versus time immersion in PBS bufferedsolution

Rugosity (nm)

LDH Ca2AlCl Zn2AlCO3 Mg2AlCO3 Mg3AlCO3

Initial LDH roughness lt025 lt025 lt025 lt025

anti-casein 0 h in PBS 082 071 102 044LDH 72 h in PBS 066 057 102 052

170 h in PBS 105 068 085 062

carbon interlayer The covered OH groups of the LDH sur-faces layer were suitable for the hydrogen bonding interac-tion with the polar function of the amino acids of antibodyThe follow-up of the surface evaluation during time in

analyzed solution shows no significant changes This isprobably due to the high affinity of the anti-bodyLDHbond that maintains stable the antibody linkage

322 Morphology Characterization

The morphology of the different biomembranes beforeand after immobilization of antibody was carried out Thefollow-up of the hybrid biomembrane stability in PBSsolution were carried out versus time the day of depo-sition after 3 days after 7 days As shown in all AFMscan images in Figure 3 the used immobilization methodpermits a good transfer of the antibody This biologicaltransfer is stable in time after 7 days In fact the rugosityof LDH surfaces is enhanced from less than 025 nm to082 nm (for Ca2Al) 045 and 080ndash102 nm (for hydro-talcites Mg3Al resp Mg2Al) and 071 nm (for Zn2Al) seeTable II After 72 hours (respectively 170 h) in PBS solu-tion no significant variations of rugosity were measuredIn fact the measured values of rugosity varied from 065ndash068 nm for Mg3AlCO3 and Zn2AlCO3 to 105 nm forCa2AlCl indicating the stable antibody linked materialsThese results are confirmed by Infra Red spectroscopyspectra In fact the follow-up of the interaction of theantibody with LDH carried out by IRTF in ATR modeshows that no significant change of spectra is observedafter 10 days of immersion in PBS The obtained bandsof spectra do not change during the follow-up study ofthe effect of the immersion in PBS solution The antibodylinked material can be considered stable for biodetectionapplication In fact after more than 15 days in solutiontest the antibody is well immobilized onto the LDH

4 CONCLUSION

In the present work we have presented a comparativestudy of different anti--caseinLDH materials The newapproach used was based on auto-assembled hybrid mate-rials The obtained immobilization of antibody onto LDH

materials can be considered highly stable in analyte solu-tion for more than 10 daysIn this study LDH materials can be considered as good

host matrices for antibody immobilization Finally thesematrices do not affect the anti-casein properties as shownthrough IRTF spectra and can be considered as a suitablebiomembrane for the casein detectionIn conclusion the feasibility of improved antibody

immobilization into LDHs materials for casein biosensorapplication is demonstrated This method will be used forfabricating the -casein biosensor and studying its perfor-mances such as sensibility selectivity and life time

Acknowledgments This work was supported by theTunisian-Spain project AECI (A971107) the Tuniso-Marocco project 06TM86 and NATO (CBPNUKRCLG982955) The technical assistance on AFM and XRDcharacterizations of respectively D Caballero from PCBBarcelona (Spain) and M Marzouki from INRAP(Tunisia) is gratefully acknowledged

References and Notes

1 S Cosnier and C Gondran Analusis 27 558 (1999)2 W Jin and J D Brennan Anal Chim Acta 461 1 (2002)3 W Hartmeier Immobilized Biocatalysts edited by W Hartmeier

Springer-Verlage Berlin (1988)4 F Kuralay H Oumlzyoumlruumlk and A Yildiz Sens Actuators B 114 500

(2006)5 Y Li W Yang Y Baim and C Sun Electroanalysis 18 499 (2006)6 S Cosnier Biosens Bioelectron 14 443 (1999)7 L Puech E Perez I Rico-Lattes M Bon and A Lattes Colloids

Surf A 167 123 (2000)8 R S Marks A Novoa D Thomassey and S Cosnier Anal

Bioanal Chem 374 1056 (2002)9 J S Thieacuteblemont J L Gabelle and M F Planche Synth Met 66

243 (1994)10 M Shinitzky F Nudelman Y Barda R Haimovitz E Chen and

D W Deamer Origins Life Evol Biosphere 32 285 (2002)11 E P Giannelis D G Nocera and T J Pinnavaia Inorg Chem 26

203 (1987)12 M Meyn K Beneke and G Lagaly Inorg Chem 29 5201 (1990)13 H Quiquampoix S Servagent-Noinville and M H Baron Enzymes

in the Environment edited by R G Burns and R P Dick MarcelDekker New York (2002) p 285

14 L Gianfreda M A Rao F Saccomandi F Sannino andA Violante Dev Soil Sci 28B 301 (2002)

15 J H Choy J Phys Chem Solids 65 373 (2004)16 A I Khan and D OrsquoHare J Mater Chem 12 3191 (2002)17 A Maaref H Barhoumi M Rammah C Martelet

N Jaffrezic-Renault C Mousty and S Cosnier Sens Actuators B123 671 (2007)

18 J V de Melo S Cosnier C Mousty C Martelet andN Jaffrezic-Renault Anal Chem 74 4037 (2002)

19 J G Miller R H Bates T A Boyer and D R Durham Porousnon-macroporous inorganic oxide carrier body for immobilizingmicroorganisms for body remediation US Patent No 6107067(2000)

20 V Ambrogi G Fardella G Grandolini and L Perioli Int J Pharm220 23 (2001)



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REVIEW


21 V Ambrogi G Fardella G Grandolini L Perioli and M C TiraltiAAPS PharmSciTech 3 77 (2002)

22 B Li J He D G Evans and X Duan Appl Clay Sci 27 199(2004)

23 M D Arco E Cebadera S Gutierrez C Martin M J MonteroV Rives J Rocha and M A Sevilla J Pharm Sci 93 1649(2004)

24 L Ren L Wan and X Duan Int J Nanotechnol 3 54(2006)

25 H Nakayama K Takeshita and M Tsuhako J Pharm Sci 922419 (2003)

26 C D Hoyo Appl Clay Sci 36 103 (2007)27 V Rives Layered Double Hydroxides Present and Future Nova

Science Publishers New York (2001) pp 285ndash32128 S Y Kwak Y J Jeong J S Park and J H Choy Solid State Ion

151 229 (2002)29 J H Choy S Y Kwak J S Park Y J Jeong and J Portier J Am

Chem Soc 121 1399 (1999)30 L Ren J He S Zhang D G Evans and X Duan J Mol Catal

B Enzym 18 3 (2002)

31 D D Borole U R Kapadi P P Mahulikar and D G HundiwaleDesigned Monomers Polym 9 1 (2006)

32 A K Ioannou A A Pantazaki S T H Girousi M C MillotC Vidal-Madjar and A N Voulgaropoulos Electroanalysis 18 456(2006)

33 R Ashaffenburg J Dairy Res 30 259 (1963)34 J Garnier B Ribadeau-Dumas and G Mocquot J Dairy Res 31

133 (1964)35 L Moio L Chianese A Di Luccia R Mauriello C Stingo

and F Addeo Electrophoresis Forum rsquo89 edited by B J RadolaMunchen (1989) pp 475ndash478

36 Z M Baccar D Caballero N Zine N Jaffrezic-Renault andA Errachid Sensors Lett 7 676 (2009)

37 I Rousselot C Taviot-Gueacuteho F Leroux P Leacuteone P Palvadeauand J P Besse J Solid State Chem 167 137 (2002)

38 J T Kloprogge D Wharton L Hickey and R L Frost AmMineral 87 623 (2002)

39 V R L Constantino and T J Pinnavaia Inorg Chem 34 883(1995)

40 Y You H T Zhao and G F Vance Appl Clay Sci 21 217 (2002)



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Z M Baccar received his PhD in Electronic devices from Ecole Centrale de Lyon Francein 1996 He was assistant professor in University of Monastir Tunisia in 1996 than inuniversity of 7 Nov in Carthage in 2000 Since 2008 he joined the National Institute ofResearch in Physics and Chemistry Analysis in Sidi Thabet as a researcher His researchactivities consist on the development and the preparation of membranes for biosensors(ISFETs ISEs ) based on hybrid nanomaterials He is leader in 4 Spanish-Tunisian(AECI and AECID) and one Morocco-Tunisian (CMPTM) program on development of sen-sors for biomedical and environmental applications

S Hidouri received his bachelor degree from University of 7 Nov of Carthage the facultyof Sciences of Bizerte (FSB) in 2004 He received the Theoretical master in Biocomputerfrom the High School of Computer Science (ENSI) in 2005 In 2009 he received a secondmaster in hydrobiology from the University of 7 Nov of Carthage the faculty of Sciencesof Bizerte (FSB) Actually he is a PhD student

N El Bari was awarded a PhD in Biology from the University Moulay Ismail (MeknesMorocco) in 1995 Since 1999 she has been a professor at the Faculty of Sciences in Meek-ness and is currently head of the Biotechnology Agroalimentary and Biomedical AnalysisGroup One of her research interest is the application of electronic nose techniques to foodspoilage

N Jaffrezic-Renault received her engineering degree from the Ecole Nationale Supeacuterieurede Chimie Paris in 1971 and the Doctorat drsquoEtat eacutes Sciences Physiques from the Uni-versity of Paris in 1976 She joined Ecole Centrale de Lyon France in 1984 and ClaudeBernard University Lyon 1 in 2007 As Director of Research at the Centre National de laRecherche Scientifique and the president of the chemical micro sensor club (CMC2) herresearch activities include the preparation and physicochemical characterization of mem-branes for chemical sensors (ISFETs optical fiber sensors electrochemical sensors) She iscurrently leading 6 PhD students 2 MSc These students are involved in one Europeanproject on microsystem for diagnostics and in several national projects for the developmentof microsystems for environmental monitoring and for food safety She is leader in a nationalprogram (INTEGREAU) involving 4 academic partners and 2 companies for the develop-

ment and industrial transfer of a generic microsystem for the monitoring of natural waters according to the frameworkdirective on water quality She published more than 350 papers with around 4000 citations



IP 16111621163Fri 16 Apr 2010 094508

REVIEW






1minusX M3+X (OH)2]

x+ middot[Axq]















IP 16111621163Fri 16 Apr 2010 094508R

EVIEW




















0 20 40 60 80

(-12

7)

(006

)

(040

)

(-12

3)

(003

)

2θ (deg)

Ca2Al

Rel

ativ

e in

tens

ity (

)

Zn2Al(116

)(0

113)

(113

)(1

10)

(011

1)

(101

0)

(018

)(1

07)(0

15)

(012

)(1

01)

(006

)

(018

)

(015

)

(012

)

(110

)(1

13)(003

)

Mg2Al

(202

6)(2

23)

(111

8)

(021

0)

(001

8)

(001

2)

(006

)

Mg3Al

(002

)

(-44

6)(2

46)

(206

)(044

)(3

23)

(400

)

(004

)




IP 16111621163Fri 16 Apr 2010 094508

REVIEW



Ca2Al


dspacing (nm)










4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

ν (cmndash1)


ν (cmndash1)

(b)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

Ant

i sym

DO

Mg3AlMg3Alab1j

Mg3Alab10j

Carboxylate 0CO-

sym

2700-3150


2650-2560


ν (cmndash1)

(c)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

CaAl

Ca2alab10jCa2alab1j

ν (cmndash1)

(d)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO





IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



(a)



(b)


Fig 3 Continued



IP 16111621163Fri 16 Apr 2010 094508

REVIEW




(c)






IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



Rugosity (nm)




170 h in PBS 105 068 085 062





4 CONCLUSION
























IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)













IP 16111621163Fri 16 Apr 2010 094508

REVIEW






1minusX M3+X (OH)2]

x+ middot[Axq]















IP 16111621163Fri 16 Apr 2010 094508R

EVIEW




















0 20 40 60 80

(-12

7)

(006

)

(040

)

(-12

3)

(003

)

2θ (deg)

Ca2Al

Rel

ativ

e in

tens

ity (

)

Zn2Al(116

)(0

113)

(113

)(1

10)

(011

1)

(101

0)

(018

)(1

07)(0

15)

(012

)(1

01)

(006

)

(018

)

(015

)

(012

)

(110

)(1

13)(003

)

Mg2Al

(202

6)(2

23)

(111

8)

(021

0)

(001

8)

(001

2)

(006

)

Mg3Al

(002

)

(-44

6)(2

46)

(206

)(044

)(3

23)

(400

)

(004

)




IP 16111621163Fri 16 Apr 2010 094508

REVIEW



Ca2Al


dspacing (nm)










4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

ν (cmndash1)


ν (cmndash1)

(b)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

Ant

i sym

DO

Mg3AlMg3Alab1j

Mg3Alab10j

Carboxylate 0CO-

sym

2700-3150


2650-2560


ν (cmndash1)

(c)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

CaAl

Ca2alab10jCa2alab1j

ν (cmndash1)

(d)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO





IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



(a)



(b)


Fig 3 Continued



IP 16111621163Fri 16 Apr 2010 094508

REVIEW




(c)






IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



Rugosity (nm)




170 h in PBS 105 068 085 062





4 CONCLUSION
























IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)













IP 16111621163Fri 16 Apr 2010 094508R

EVIEW




















0 20 40 60 80

(-12

7)

(006

)

(040

)

(-12

3)

(003

)

2θ (deg)

Ca2Al

Rel

ativ

e in

tens

ity (

)

Zn2Al(116

)(0

113)

(113

)(1

10)

(011

1)

(101

0)

(018

)(1

07)(0

15)

(012

)(1

01)

(006

)

(018

)

(015

)

(012

)

(110

)(1

13)(003

)

Mg2Al

(202

6)(2

23)

(111

8)

(021

0)

(001

8)

(001

2)

(006

)

Mg3Al

(002

)

(-44

6)(2

46)

(206

)(044

)(3

23)

(400

)

(004

)




IP 16111621163Fri 16 Apr 2010 094508

REVIEW



Ca2Al


dspacing (nm)










4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

ν (cmndash1)


ν (cmndash1)

(b)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

Ant

i sym

DO

Mg3AlMg3Alab1j

Mg3Alab10j

Carboxylate 0CO-

sym

2700-3150


2650-2560


ν (cmndash1)

(c)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

CaAl

Ca2alab10jCa2alab1j

ν (cmndash1)

(d)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO





IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



(a)



(b)


Fig 3 Continued



IP 16111621163Fri 16 Apr 2010 094508

REVIEW




(c)






IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



Rugosity (nm)




170 h in PBS 105 068 085 062





4 CONCLUSION
























IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)













IP 16111621163Fri 16 Apr 2010 094508

REVIEW



Ca2Al


dspacing (nm)










4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

ν (cmndash1)


ν (cmndash1)

(b)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

Ant

i sym

DO

Mg3AlMg3Alab1j

Mg3Alab10j

Carboxylate 0CO-

sym

2700-3150


2650-2560


ν (cmndash1)

(c)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO

CaAl

Ca2alab10jCa2alab1j

ν (cmndash1)

(d)

4000 3500 3000 2500 2000 1500 1000 500

000

005

010

015

020

025

DO





IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



(a)



(b)


Fig 3 Continued



IP 16111621163Fri 16 Apr 2010 094508

REVIEW




(c)






IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



Rugosity (nm)




170 h in PBS 105 068 085 062





4 CONCLUSION
























IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)













IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



(a)



(b)


Fig 3 Continued



IP 16111621163Fri 16 Apr 2010 094508

REVIEW




(c)






IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



Rugosity (nm)




170 h in PBS 105 068 085 062





4 CONCLUSION
























IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)













IP 16111621163Fri 16 Apr 2010 094508

REVIEW




(c)






IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



Rugosity (nm)




170 h in PBS 105 068 085 062





4 CONCLUSION
























IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)













IP 16111621163Fri 16 Apr 2010 094508R

EVIEW



Rugosity (nm)




170 h in PBS 105 068 085 062





4 CONCLUSION
























IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)













IP 16111621163Fri 16 Apr 2010 094508

REVIEW











B Enzym 18 3 (2002)












Stable Immobilization of Anti-Beta Casein Antibody onto Layered Double Hydroxides Materials for...

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Transcript of Stable Immobilization of Anti-Beta Casein Antibody onto Layered Double Hydroxides Materials for...