Skip to main content
SpringerLink
Log in

Layer-by-layer immobilization of horseradish peroxidase on a gold electrode modified with colloidal gold nanoparticles

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract.

An amperometric biosensor, based on layer-by-layer self-assembly of colloidal gold nanoparticles, cysteine and horseradish peroxidase on Nafion modified electrode surface by electrostatic adsorption, has been used for the determination of hydrogen peroxide. The electrochemical behavior of the multilayer film was studied by cyclic voltammetry, linear sweep voltammetry and chronoamperometry. The step layer-by-layer adsorption interface morphology was further characterized by means of electrochemical impedance spectroscopy and cyclic voltammetry. The performance and factors influencing the resulted biosensor were studied in detail. The sensor displayed an excellent electrocatalytic response to the reduction of H2O2 without the aid of an electron transfer mediator. Linear response to H2O2 was obtained for the concentration range from 1.6 µM to 2.4 mM under optimized conditions. The detection limit of the biosensor was 0.5 µM (S/N = 3), and the sensor achieved 95% of the steady-state current within 10 s. The sensor exhibited high sensitivity, selectivity and stability.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • G Decher J D Hong (1991) ArticleTitleBuildup of ultrathin multilayer films by a self-assembly process, consecutive adsorption of anionic and cationic bipolar amphiphiles Makromol Chem Macromol Symp 46 321 Occurrence Handle1:CAS:528:DyaK3MXms1Citbs%3D

    CAS  Google Scholar 

  • G Decher J D Hong (1991) ArticleTitleBuildup of ultrathin multilayer films by a self-assembly process. II. Consecutive adsorption of anionic and cationic bipolar amphiphiles and polyelectrolytes on charged surfaces Ber Bunsen-Ges Phys Chem 95 1430 Occurrence Handle1:CAS:528:DyaK38XmvVajtA%3D%3D

    CAS  Google Scholar 

  • G Decher (1997) ArticleTitleFuzzy nanoassemblies: toward layered polymeric multicomposites Science 277 1232 Occurrence Handle10.1126/science.277.5330.1232 Occurrence Handle1:CAS:528:DyaK2sXlslaitL4%3D

    Article  CAS  Google Scholar 

  • Y Lvov K Ariga I Ichinose (1995) ArticleTitleAssembly of multicomponent protein films by means of elelctrostatic layer-by-layer adsorption J Am Chem Soc 117 6117 Occurrence Handle10.1021/ja00127a026 Occurrence Handle1:CAS:528:DyaK2MXlvFWkt7g%3D

    Article  CAS  Google Scholar 

  • Y Lvov G Decher G Sukhorukov (1993) ArticleTitleAssembly of thin films by means of successive deposition of alternate layers of DNA and poly(allylamine) Macromolecules 26 5396 Occurrence Handle10.1021/ma00072a016 Occurrence Handle1:CAS:528:DyaK3sXls1Wrtr0%3D

    Article  CAS  Google Scholar 

  • S A Miscoria J Desbrieres G D Barrera P Labbé G A Rivas (2006) ArticleTitleGlucose biosensor based on the layer-by-layer self-assembling of glucose oxidase and chitosan derivatives on a thiolated gold surface Anal Chim Acta 578 137 Occurrence Handle10.1016/j.aca.2006.06.060 Occurrence Handle1:CAS:528:DC%2BD28XhtVSitrvE

    Article  CAS  Google Scholar 

  • J F Rusling (2000) NoChapterTitle Y Lvov H Mohwald (Eds) Protein architecture: interfacing molecular assemblies and immobilization biotechnology Dekker New York 337

    Google Scholar 

  • Z Li N F Hu (2003) ArticleTitleDirect electrochemistry of heme proteins in their layer-by-layer films with clay nanoparticles J Electroanal Chem 558 155 Occurrence Handle10.1016/S0022-0728(03)00390-5 Occurrence Handle1:CAS:528:DC%2BD3sXotlylsrk%3D

    Article  CAS  Google Scholar 

  • H Y Ma N F Hu J F Rusling (2000) ArticleTitleElectroactive myoglobin films grown layer-by-layer with poly(styrenesulfonate) on pyrolytic graphite electrodes Langmuir 16 4969 Occurrence Handle10.1021/la991296t Occurrence Handle1:CAS:528:DC%2BD3cXisFGhsbk%3D

    Article  CAS  Google Scholar 

  • Y Lvov F Caruso (2001) ArticleTitleBiocolloids with ordered urease multilayer shells as enzymatic reactors Anal Chem 73 4212 Occurrence Handle10.1021/ac010118d Occurrence Handle1:CAS:528:DC%2BD3MXls12qtL8%3D

    Article  CAS  Google Scholar 

  • S X Zhang W W Yang Y M Niu Y C Li M Zhang C Q Sun (2006) ArticleTitleConstruction of glucose biosensor based on sorption of glucose oxidase onto multilayers of polyelectrolyte/nanoparticles Anal Bioanal Chem 384 736 Occurrence Handle10.1007/s00216-005-0190-7 Occurrence Handle1:CAS:528:DC%2BD28Xhtl2msrs%3D

    Article  CAS  Google Scholar 

  • L Y Zhao H Y Liu N F Hu (2006) ArticleTitleAssembly of layer-by-layer films of heme proteins and single-walled carbon nanotubes: electrochemistry and elelctrocatalysis Anal Bioanal Chem 384 414 Occurrence Handle10.1007/s00216-005-0204-5 Occurrence Handle1:CAS:528:DC%2BD28XhsF2gsA%3D%3D

    Article  CAS  Google Scholar 

  • H B Shi Y Yang J D Huang Z X Zhao X H Xu J Anzai T Osa Q Chen (2006) ArticleTitleAmperometric choline biosensors prepared by layer-by-layer deposition of choline oxidase on the Prussian blue-modified platinum elelctrode Talanta 70 852 Occurrence Handle10.1016/j.talanta.2006.02.012 Occurrence Handle1:CAS:528:DC%2BD28XhtVWqt7vM

    Article  CAS  Google Scholar 

  • Y D Jin Y Shao S J Dong (2003) ArticleTitleDirect electrochemistry and surface plasmon resonance characterization of alternate layer-by-layer self-assembled DNA-myolobin thin films on chemically modified gold surfaces Langmuir 19 4771 Occurrence Handle10.1021/la026873j Occurrence Handle1:CAS:528:DC%2BD3sXjt1SgsLg%3D

    Article  CAS  Google Scholar 

  • L Q Luo J Y Liu Z X Wang X R Yang S J Dong E K Wang (2001) ArticleTitleFabrication of layer-by-layer deposited multilayer films containing DNA and its interaction with methyl green Biophys Chem 94 11 Occurrence Handle10.1016/S0301-4622(01)00204-6 Occurrence Handle1:CAS:528:DC%2BD3MXptVSltLo%3D

    Article  CAS  Google Scholar 

  • R Pei X Cui X Yang E Wang (2001) ArticleTitleAssembly of alternating polycation and DNA multilayer films by electrostatic layer-by-layer adsorption Biomacromol 2 463 Occurrence Handle10.1021/bm0001289 Occurrence Handle1:CAS:528:DC%2BD3MXhs1agtb8%3D

    Article  CAS  Google Scholar 

  • Y L Zhou Z Li N F Hu Y H Zeng J F Rusling (2002) ArticleTitleLayer-by-layer assembly of ultrathin films of hemoglobin and clay nanoparticles with electrochemical and catalytic activity Langmuir 18 8573 Occurrence Handle10.1021/la026120x Occurrence Handle1:CAS:528:DC%2BD38Xntlaiurw%3D

    Article  CAS  Google Scholar 

  • M H Yang Y Yang H F Yang G L Shen R Q Yu (2006) ArticleTitleLayer-by-layer self-assembled multilayer films of carbon nanotubes and platinum nanoparticles with polyelectrolyte for the fabrication of biosensors Biomaterials 27 246 Occurrence Handle10.1016/j.biomaterials.2005.05.077 Occurrence Handle1:CAS:528:DC%2BD2MXhtVShtbzL

    Article  CAS  Google Scholar 

  • E Topoglidis Y Astuti F Duriaux M GrStzel J R Durrant (2003) ArticleTitleDirect electrochemistry and nitric oxide interaction of heme proteins adsorbed on nanocrystalline tin oxide electrodes Langmuir 19 6894 Occurrence Handle10.1021/la034466h Occurrence Handle1:CAS:528:DC%2BD3sXltV2is7g%3D

    Article  CAS  Google Scholar 

  • Y Y Sun F Yan W S Yang C Q Sun (2006) ArticleTitleMultilayered construction of glucose oxidase and silica nanoparticles on Au electrodes based on layer-by-layer covalent attachment Biomatericals 27 4042 Occurrence Handle10.1016/j.biomaterials.2006.03.014 Occurrence Handle1:CAS:528:DC%2BD28Xjslyitb8%3D

    Article  CAS  Google Scholar 

  • C Tedeschi F Caruso H Mohwald S Kirstein (2000) ArticleTitleAdsorption and desorption behavior of an anionic pyrene chromophore in sequentially deposited polyelectrolyte-dye thin films J Am Chem Soc 122 5841 Occurrence Handle10.1021/ja994029i Occurrence Handle1:CAS:528:DC%2BD3cXjsF2qtLY%3D

    Article  CAS  Google Scholar 

  • K Ariga Y Lvov T Kunitake (1997) ArticleTitleAssembling alternate dye–polyion molecular films by electrostatic layer-by-layer adsorption J Am Chem Soc 119 2224 Occurrence Handle10.1021/ja963442c Occurrence Handle1:CAS:528:DyaK2sXht1OhsL0%3D

    Article  CAS  Google Scholar 

  • X D Chen J Lang M H Liu (2002) ArticleTitleLayer-by-layer assembly DNA-dye complex films Thin Solid Films 409 227 Occurrence Handle1:CAS:528:DC%2BD38XjslCltr0%3D

    CAS  Google Scholar 

  • J D Huang Y Yang H B Shi Z Song Z X Zhao J Anzai T Osa Q Chen (2006) ArticleTitleMulti-walled carbon nanotubes-based glucose biosensor prepared by a layer-by-layer technique Mater Sci Eng C 26 113 Occurrence Handle10.1016/j.msec.2005.06.002 Occurrence Handle1:CAS:528:DC%2BD2MXhtlCns7%2FF

    Article  CAS  Google Scholar 

  • X B Yan X J Chen B K Tay K A Khor (2007) ArticleTitleTransparent and flexible glucose biosensor via layer-by-layer assembly of multi-wall carbon nanotubes and glucose oxidase Electrochem Comm 9 1269 Occurrence Handle10.1016/j.elecom.2006.12.022 Occurrence Handle1:CAS:528:DC%2BD2sXlslGit7Y%3D

    Article  CAS  Google Scholar 

  • H T Zhao H X Ju (2006) ArticleTitleMultilayer membranes for glucose biosensing via layer-by-layer assembly of multiwall carbon nanotubes and glucose oxidase Anal Biochem 350 138 Occurrence Handle10.1016/j.ab.2005.11.034 Occurrence Handle1:CAS:528:DC%2BD28XhsF2is7o%3D

    Article  CAS  Google Scholar 

  • B Y Wu S H Hou F Yin J Li Z X Zhao (2007) ArticleTitleAmperometric glucose biosensor based on layer-by-layer assembly of multilayer films composed of chitosan, gold nanoparticles and glucose oxidase modified Pt electrode Biosens Bioelectron 22 838 Occurrence Handle10.1016/j.bios.2006.03.009 Occurrence Handle1:CAS:528:DC%2BD28XhtlalsbzJ

    Article  CAS  Google Scholar 

  • M A Correa-Duarte M Gierig N A Kotov L M Liz-Marzan (1998) ArticleTitleControl of packing order of self-assembled monolayers of magnetite nanoparticles with and without SiO2 coating by microwave irradiation Langmuir 14 6430 Occurrence Handle10.1021/la9805342 Occurrence Handle1:CAS:528:DyaK1cXmt1yhtrk%3D

    Article  CAS  Google Scholar 

  • W W Yang Y C Li Y Bai C Q Sun (2006) ArticleTitleHydrogen peroxide biosensor based on mylglobin/colloidal gold nanoparticles immobilized on glassy carbon electrode by a Nafion film Sens Actuators B 115 42 Occurrence Handle10.1016/j.snb.2005.08.017

    Article  Google Scholar 

  • Y Zhuo R Yuan Y Q Chai Y Zhang X L Li N Wang Q Zhu (2006) ArticleTitleAmperometric enzyme immunosensors based on layer-by-layer assembly of gold nanoparticles and thionine on Nafion modified electrode surface for α-1-fetoprotein determinations Sens Actuators B 114 631 Occurrence Handle10.1016/j.snb.2005.04.051

    Article  Google Scholar 

  • A L Crumbliss S C Perine J Stonehuerner K R Twbergen J Zhao R W Henkens J P O‘Daly (1992) ArticleTitleColloidal gold as a biocompatible immobilization matrix suitable for the fabrication of enzyme electrodes by electrodeposition Biotechnol Bioeng 40 483 Occurrence Handle10.1002/bit.260400406 Occurrence Handle1:CAS:528:DyaK38XlsVKnsrY%3D

    Article  CAS  Google Scholar 

  • Y Xiao H X Ju H Y Chen (1999) ArticleTitleHydrogen peroxide sensor based on horseradish peroxidase-labeled Au colloids immobilized on gold electrode surface by cysteamine monolayer Anal Chim Acta 391 73 Occurrence Handle10.1016/S0003-2670(99)00196-8 Occurrence Handle1:CAS:528:DyaK1MXjtFKitr8%3D

    Article  CAS  Google Scholar 

  • L Wang E Wang (2004) ArticleTitleA novel hydrogen peroxide sensor based on horseradish peroxidase immobilized on colloidal Au modified ITO electrode Electrochem Comm 6 225 Occurrence Handle10.1016/j.elecom.2003.12.004 Occurrence Handle1:CAS:528:DC%2BD2cXksFaltQ%3D%3D

    Article  CAS  Google Scholar 

  • F Patolsky T Gabriel I Willner (1999) ArticleTitleControlled electrocatalysis by microperoxidase-11 and Au-nanoparticle superstructures on conductive supports J Electroanal Chem 479 69 Occurrence Handle10.1016/S0022-0728(99)00426-X Occurrence Handle1:CAS:528:DyaK1MXotVGntb4%3D

    Article  CAS  Google Scholar 

  • S H Chen R Yuan Y Q Chai L Xu N Wang X L Li L Y Zhang (2006) ArticleTitleAmperometric hydrogen peroxide biosensors based on the immobilization of horseradish (HRP) on the layer-by-layer assembly films of gold colloidal nanoparticles and toluidine blue Electroanalysis 17 471 Occurrence Handle10.1002/elan.200503424

    Article  Google Scholar 

  • J B Jia B Q Wang A Wu G J Cheng Z Li S J Dong (2002) ArticleTitleA method to construct a third-generation horseradish peroxidase biosensor: self-assembling gold nanoparticles to three-dimensional sol-gel network Anal Chem 74 2217 Occurrence Handle10.1021/ac011116w Occurrence Handle1:CAS:528:DC%2BD38XitlCjur0%3D

    Article  CAS  Google Scholar 

  • C X Lei S Q Hu G L Shen R Q Yu (2003) ArticleTitleImmobilization of horseradish peroxidase to a nano-Au monolayer modified chitosan-entrapped carbon paste electrode for the detection of hydrogen peroxide Talanta 59 981 Occurrence Handle10.1016/S0039-9140(02)00641-0 Occurrence Handle1:CAS:528:DC%2BD3sXis1ejurw%3D

    Article  CAS  Google Scholar 

  • Y T Kong M Boopathi Y B Shim (2003) ArticleTitleDirect electrochemistry of horseradish peroxidase bonded on a conducting polymer modified glassy carbon electrode Biosensors Bioelectronics 19 227 Occurrence Handle10.1016/S0956-5663(03)00216-1 Occurrence Handle1:CAS:528:DC%2BD3sXovVWltbw%3D

    Article  CAS  Google Scholar 

  • Y Liu R Yuan Y Q Chai D P Tang J Y Dai X Zhong (2006) ArticleTitleDirect electrochemistry of horseradish peroxidase immobilized on gold colloid/cysteine/nafion-modified platinum disk electrode Sens Actuators B 115 109 Occurrence Handle10.1016/j.snb.2005.08.048

    Article  Google Scholar 

  • H Cai C Xu P He Y Fang (2001) ArticleTitleColloid Au-enhanced DNA immobilization for the electrochemical detection of sequencespecific DNA J Electroanal Chem 510 78 Occurrence Handle10.1016/S0022-0728(01)00548-4 Occurrence Handle1:CAS:528:DC%2BD3MXlsFKisbw%3D

    Article  CAS  Google Scholar 

  • R Yuan Y Zhuo Y Q Chai Y Zhang A L Sun (2007) ArticleTitleDetermination of carcinoebryonic antigen using a novel amperometric enzyme-electrode based on layer-by-layer assembly of gold nanoparticles and thionine Sci China SerB-Chem 50 97 Occurrence Handle10.1007/s11426-007-0017-9 Occurrence Handle1:CAS:528:DC%2BD2sXhvFSgu78%3D

    Article  CAS  Google Scholar 

  • Y Tian M Shioda S Kasahara T Okajima L Mao T Hisabori T Ohsaka (2002) ArticleTitleA facilitated electron transfer of copper-zinc superoxide dismutase (SOD) based on cysteine-bridged SOD electrode Biochim Biophy Acta 1569 151 Occurrence Handle1:CAS:528:DC%2BD38Xjt1KgsL4%3D

    CAS  Google Scholar 

  • Y Xiao H X Ju H Y Chen (1999) ArticleTitleHydrogen peroxide sensor based on horseradish peroxidase-labeled Au colloids immobilized on gold electrode surface by cysteamine monolayer Anal Chim Acta 391 73 Occurrence Handle10.1016/S0003-2670(99)00196-8 Occurrence Handle1:CAS:528:DyaK1MXjtFKitr8%3D

    Article  CAS  Google Scholar 

  • S Q Liu H X Ju (2003) ArticleTitleReagentless glucose biosensor based on direct electron transfer of glucose oxidase immobilized on colloidal gold modified carbon paste electrode Biosens Bioelectron 19 177 Occurrence Handle10.1016/S0956-5663(03)00172-6 Occurrence Handle1:CAS:528:DC%2BD3sXovVWltLg%3D

    Article  CAS  Google Scholar 

  • J J Feng J J Xu H Y Chen (2007) ArticleTitleDirect electron transfer and electrocatalysis of hemoglobin adsorbed on mesoporous carbon through layer-by-layer assembly Biosen Bioelectron 22 1618 Occurrence Handle10.1016/j.bios.2006.07.022 Occurrence Handle1:CAS:528:DC%2BD2sXhtlOmsr4%3D

    Article  CAS  Google Scholar 

  • L Meites (1965) Polarographic techiques EditionNumber2 Wiley New York

    Google Scholar 

  • A M Bond (1980) Modern polagraphic methods in analytical chemistry Marcel Dekker New York

    Google Scholar 

  • L Qiao X Q Yang (2006) ArticleTitleComposite film of carbon nanotubes and chitosan for preparation of amperometric hydrogen peroxide biosensor Talanta 68 721 Occurrence Handle10.1016/j.talanta.2005.05.030

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. College of Chemistry, Chongqing Normal University, Chongqing, P.R. China

    Yan Liu, Tongmou Geng & Jie Gao

Authors
  1. Yan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tongmou Geng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jie Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yan Liu.

Additional information

Correspondence: Yan Liu, College of Chemistry, Chongqing Normal University, Chongqing 400047, P.R. China

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, Y., Geng, T. & Gao, J. Layer-by-layer immobilization of horseradish peroxidase on a gold electrode modified with colloidal gold nanoparticles. Microchim Acta 161, 241–248 (2008). https://doi.org/10.1007/s00604-007-0779-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00604-007-0779-y

Search

Navigation