Phospholipid monolayer coated microfabricated electrodes to model the interaction of molecules with biomembranes

Abstract The hanging mercury (Hg) drop electrode (HMDE) has a classical application as a tool to study adsorption and desorption processes of surface organic films due to its: (a) atomically smooth surface and, (b) hydrophobicity at its potential of zero charge. In this study we report on a replacement of the HMDE for studying supported organic layers in the form of platinum (Pt) working electrodes fabricated using lithography techniques on which a thin film of Hg is electrodeposited. These wafer-based Pt/Hg electrodes are characterised and compared to the HMDE using rapid cyclic voltammetry (RCV) and show similar capacitance–potential profiles while being far more mechanically stable and consuming considerably less Hg over their lifetime of several months. The electrodes have been used to support self-assembled phospholipid monolayers which are dynamic surface coatings with unique dielectric properties. The issue of surface contamination has been solved by regenerating the electrode surface prior to phospholipid coating by application of extreme cathodic potentials more negative than −2.6 V (vs. Ag/AgCl). The phospholipid coated electrodes presented in this paper mimic one half of a phospholipid bilayer and exhibit interactions with the biomembrane active drug molecules chlorpromazine, and quinidine. The magnitudes of these interactions have been assessed by recording changes in the capacitance–potential profiles in real time using RCV at 40 V s −1 over potential ranges >1 V. A method for electrode coating with phospholipids with the electrodes fitted in a flow cell device has been developed. This has enabled sequential rapid cleaning/coating/interaction cycles for the purposes of drug screening and/or on-line monitoring for molecules of interest.

[1]  F. Leermakers,et al.  Substrate-induced structural changes in electrode-adsorbed lipid layers: Experimental evidence from the behaviour of phospholipid layers on the mercury-water interface , 1990 .

[2]  R. Ofoli,et al.  Comparison of liposomes formed by sonication and extrusion: rotational and translational diffusion of an embedded chromophore. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[3]  D. Chapman,et al.  LI. A contribution to the theory of electrocapillarity , 1913 .

[4]  A. Nelson Electrochemical analysis of a phospholipid phase transition , 2007 .

[5]  F. Scholz,et al.  The lipid composition determines the kinetics of adhesion and spreading of liposomes on mercury electrodes. , 2008, Bioelectrochemistry.

[6]  V. Vetterl,et al.  Application of carbon electrodes modified with a mercury layer of a different thickness for studies of the adsorption and kinetics of phase transients of cytidine , 2002 .

[7]  F. Leermakers,et al.  Substrate-induced structural changes in electrode-adsorbed lipid layers: A self-consistent field theory , 1990 .

[8]  A. Schmidtchen,et al.  An electrochemical study into the interaction between complement-derived peptides and DOPC mono- and bilayers. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[9]  J. Buffle,et al.  Deposition and Stripping Properties of Mercury on Iridium Electrodes , 1986 .

[10]  S. Babkina,et al.  Amperometric biosensor based on denatured DNA for the study of heavy metals complexing with DNA and their determination in biological, water and food samples. , 2004, Bioelectrochemistry.

[11]  A. Nelson Penetration of mercury-adsorbed phospholipid monolayers by polynuclear aromatic hydrocarbons , 1987 .

[12]  S. Kounaves,et al.  Analytical Utility of the Iridium-Based Mercury Ultramicroelectrode with Square-Wave Anodic Stripping Voltammetry , 1993 .

[13]  D. Grahame The electrical double layer and the theory of electrocapillarity. , 1947, Chemical reviews.

[14]  D. Grahame On the Determination of the Differential Capacity at a Dropping Mercury Electrode , 1957 .

[15]  A. Nelson,et al.  Continuing Electrochemical Studies of Phospholipid Monolayers of Dioleoyl Phosphatidylcholine at the Mercury−Electrolyte Interface , 1998 .

[16]  Walter Schmitt,et al.  A physiological model for the estimation of the fraction dose absorbed in humans. , 2004, Journal of medicinal chemistry.

[17]  Samuel P. Kounaves,et al.  Microfabricated Array of Iridium Microdisks as a Substrate for Direct Determination of Cu2+ or Hg2+ Using Square-Wave Anodic Stripping Voltammetry , 1999 .

[18]  R. Silvennoinen,et al.  On the adsorption and kinetics of phase transients of adenosine at the different carbon electrodes modified with a mercury layer , 2003 .

[19]  J. Buffle,et al.  An iridium-based mercury-film electrode: Part I. Selection of substrate and preparation , 1987 .

[20]  R. S. Nicholson,et al.  Theory of Stationary Electrode Polarography. Single Scan and Cyclic Methods Applied to Reversible, Irreversible, and Kinetic Systems. , 1964 .

[21]  A. Nelson,et al.  Phospholipid Monolayers At The Mercury Water Interface , 1986 .

[22]  E. Disalvo,et al.  Lipid monolayers on Hg as a valid experimental model for lipid membranes under electrical fields. , 2006, Chemistry and physics of lipids.

[23]  M. Pásek,et al.  Quantification of t-tubule area and protein distribution in rat cardiac ventricular myocytes. , 2008, Progress in biophysics and molecular biology.

[24]  Emily A Hueske,et al.  Scanning electrochemical microscopy. 48. Hg/Pt hemispherical ultramicroelectrodes: fabrication and characterization. , 2003, Analytical chemistry.

[25]  D. Mandler,et al.  Characterization of n-alkanethiol self-assembled monolayers on mercury by impedance spectroscopy and potentiometric measurements , 2006 .

[26]  A Nelson,et al.  Interaction of hydrophobic organic compounds with mercury adsorbed dioleoylphosphatidylcholine monolayers. , 1990, Biochimica et biophysica acta.

[27]  A. Nelson,et al.  Initial applications of phospholipid-coated mercury electrodes to the determination of polynuclear aromatic hydrocarbons and other organic micropollutants in aqueous systems , 1988 .