Electrochemical immunosensor for the milk allergen Β-lactoglobulin based on electrografting of organic film on graphene modified screen-printed carbon electrodes

Shimaa Eissa, Chaker Tlili, Lamia L'Hocine, Mohammed Zourob

Research output: Contribution to journalArticlepeer-review

127 Scopus citations

Abstract

A novel label-free voltammetric immunosensor for sensitive detection of Β-lactoglobulin using graphene modified screen printed electrodes has been developed. The derivatization of the graphene electrode surface was achieved by electrochemical reduction of in situ generated 4-nitrophenyl diazonium cations in aqueous acidic solution, followed by electrochemical reduction of the terminal nitro groups to amines. The electrochemical modification protocol was optimized in order to generate monolayer of nitrophenyl groups on the graphene surface without complete passivation of the electrode. Unlike the reported method for graphene functionalization, we demonstrated here the ability of the electrografting of aryl diazonium salt to attach an organic film to the graphene surface in a controlled manner by choosing the suitable grafting protocol. Next, the amine groups on the graphene surface were activated using glutaraldehyde and used for the covalent immobilization of Β-lactoglobulin antibodies. Cyclic and differential pulse voltammetry carried out in an aqueous solution containing [Fe(CN)6]3-/4- redox pair have been used for the immunosensor characterization. The results demonstrated that the DPV reduction peak current of [Fe(CN)6]3-/4- decreased linearly with increasing the concentration of Β-lactoglobulin due to the formation of antibody-antigen complex on the modified electrode surface. The immunosensor obtained using this novel approach enabled a detection limit of 0.85pgmL-1 and a dynamic range from 1pgmL-1 to 100ngmL-1 of Β-lactoglobulin in PBS buffer. In addition, the immunosensor evaluated in different samples including cake, cheese snacks, a sweet biscuit, showing excellent correlation with the results obtained from commercially enzyme-linked immunosorbent assay (ELISA) method.

Original languageBritish English
Pages (from-to)308-313
Number of pages6
JournalBiosensors and Bioelectronics
Volume38
Issue number1
DOIs
StatePublished - Oct 2012

Keywords

  • Β-lactoglobulin
  • Electrochemical
  • Electrografting
  • Graphene
  • Immunosensor

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