Electrochemical Detection of Biotin Using an Interaction between Avidin and Biotin Labeled with Ferrocene at a Perfluorosulfonated Ionomer Modified Electrode

Abstract

The electrochemical behavior of biotin labeled with ferrocene was investigated at a perfluorosulfonated ionomer (Nafion ® ) film-coated glassy carbon electrode (GCE). The electrode response of the labeled biotin (LB) was improved by about fifty-fold in sensitivity by using a Nafion ® -coated GCE, compared with a plain GCE. It was found by a solid-phase avidin-biotin binding assay that the labeling with ferrocene hardly affected the affinity of a biotin moiety in the labeled biotin to avidin. The avidin-biotin interaction was estimated from the change of the electrode response caused by the formation of a complex with avidin and the labeled biotin. Furthermore, the detection of biotin was performed by a competitive reaction between the labeled biotin and unlabeled biotin for the limited binding sites of avidin.

Electrochemical Evaluation of the Interaction between Endocrine Disrupter Chemicals and Estrogen Receptor Using 17β-Estradiol Labeled with Daunomycin

“…Although electrochemical methods have many advantages (for example, they are relatively simple, rapid, inexpensive, and precise), a screening method of EDCs by electrochemical techniques has not yet been reported. We reported earlier on some simple electrochemical assays to detect the protein−ligand interaction, such as avidin−biotin − and lectin−sugar. , These methods do not require a separation procedure, since the protein−ligand interaction can be evaluated solely on the basis of changes in the electrode response of free ligand labeled with an electroactive compound. A similar attempt has been made in an immunoassay by using an antigen labeled with an electrochemically active substance for signal generation.…”

Multiplexed Assay for Proteins Based on Sequestration Electrochemistry Using the Protein Binding Electroactive Magnetic Microbeads

“…The current peak at 290 mV is due to ferrocene moiety. 15 The peak at -320 mV might be from thionine moiety. However, it is shifted slightly negatively compared to that in the previous report.…”

“…Therefore, it involves multiple reaction steps which require the addition of substrate and onerous washing/rinsing procedures. On the other hand, the use of an electroactive compound as a label for binding assay is the subject of continued research and development, [10][11][12][13][14][15][16][17] especially considering the advantages of not requiring any reagents such as substrate for enzyme reaction and being a one-step reaction, i.e. the reaction for target protein.…”

Synthetic Methodology for Asymmetric Ferrocene Derived Bio-conjugate Systems via Solid Phase Resin-based Methodology

“…[32][33][34][35][36][37][38]43 Systems targeting larger species such as biomolecules have utilized the attachment of large antibodies or immunoglobulins to ferrocene derivatives that have been embedded onto an electrochemical surface. 1,3,39,44 In each case, the potential and current intensity of the Fe III /Fe II redox couple was altered upon molecular coupling, thus producing a new spectroscopic handle indicating the presence of the analyte molecule. This change arises from the extensive overlap that occurs between the pi-system of the cyclopentadienyl rings and the iron d-orbitals.…”