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Membrane-Based Biosensing

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A special issue of Sensors (ISSN 1424-8220). This special issue belongs to the section "Biosensors".

Deadline for manuscript submissions: closed (30 September 2018) | Viewed by 19072

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Prof. Dr. Dimitrios P. Nikolelis

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Laboratory of Environmental Chemistry, Department of Chemistry, University of Athens, Panepistimiopolis-Kouponia, 15771 Athens, Greece
Interests: biosensors; analytical and environmental chemistry; nanotechnology
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Dimitrios P. Nikolelis

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Dear Colleagues,

Molecular self-assembly mimics natural systems and is a key link between physics, chemistry and biology. Molecular self-assembly can be used to create novel structures, materials, and devices for use in biosensors. Of all the self-assembled structures, thin lipid films and liposomes are the ones attracting the most attention in relation to biosensors. Like a cell membrane, lipid films and liposomes are composed of phospholipids or other amphiphiles. Their hydrophilic/hydrophobic characteristics allow them to spontaneously form organized structures.

The supported bilayer lipid membrane (BLM) provides a natural environment for embedding proteins, receptors, membrane/tissue fragments, and entire cells under nondenaturing conditions and in a well-defined orientation. This makes BLMs especially attractive for use in biosensors.

A successful biomimetically engineered device based on BLMs was the ion channel switch biosensor has been reported in the literature. The basis of this 1.5 nm nanomachine was a self-assembled artificial membrane packed with gramicidin. Ion channels were formed in the membrane by two gramicidin molecules: One in the lower layer of the membrane attached to a gold electrode and one in the upper layer tethered to biological receptors, such as antibodies or nucleotides. The detection mechanism operated by binding the target molecule to the receptor and thereby altering the population of conduction ion channel pairs within the tethered membrane. This resulted in a change in the membrane conduction. The device was capable of detecting picomolar concentrations of proteins.

Unlike planer BLMs, liposomes are microscopic, fluid-filled, pouches with endless walls that are made of layers of phospholipids identical to the phospholipids that make up cell membranes. Liposomes are typically used as the supporting substrate for immobilizing the biorecognition molecules. Liposomes are also used to amplify the optical, sound wave, and electrochemical signals.

Lipid film coated electrodes represent unique tool for preparation of enzyme biosensors and for study the mechanisms of enzymatic reactions at the surfaces. The enzyme can be incorporated into the lipid films by means of dissolution of enzyme molecules in lipid solution from which the film is prepared or by immobilization of the enzyme at the lipid film surface. Modification of supported lipid membranes and liposomes with the receptors allows substantial amplification of the signal in detection of target species using various modes of detection, such as electrochemical, optical and acoustic waves.

Free standing BLMs and supported BLMs
Metal supported sBLMs
sBLMs formed on a surface of glassy carbon, mica, metal oxide surfaces and on a ultrafiltration membranes
Bilayers stabilized by polymerization and formed on semiconductor and carbon nanotube surfaces
Polymer-supported bilayer lipid membranes
Lipid films supported on carbon nanotubes
sBLMs formed on nanomaterials
Biosensors based on supported lipid films and their analytical applications
Ion-selective sBLM and water permeability channels.
Enzyme biosensors
DNA biosensors
Affinity biosensors based on artificial and natural receptors

Prof. Dr. Dimitrios P. Nikolelis
Dr. Georgia-Paraskevi Nikoleli
Guest Editors

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Keywords

lipid membranes
biosensors
nanotechnology
liposomes
biochemical processes
biomembranes
ion channels
enzymes
DNA, receptors

Published Papers (5 papers)

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Research

14 pages, 4888 KiB

Open AccessArticle

Label-Free and Redox Markers-Based Electrochemical Aptasensors for Aflatoxin M1 Detection

by Stefanos Karapetis, Dimitrios Nikolelis and Tibor Hianik

Sensors 2018, 18(12), 4218; https://doi.org/10.3390/s18124218 - 01 Dec 2018

Cited by 31 | Viewed by 4054

Abstract

We performed a comparative analysis of the sensitivity of aptamer-based biosensors for detection mycotoxin aflatoxin M₁ (AFM1) depending on the method of immobilization of DNA aptamers and method of the detection. Label-free electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) for ferrocene labeled neutravidin layers were used for this purpose. Amino-modified DNA aptamers have been immobilized at the surface of polyamidoamine dendrimers (PAMAM) of fourth generation (G4) or biotin-modified aptamers were immobilized at the neutravidin layer chemisorbed at gold surface. In the first case the limit of detection (LOD) has been determined as 8.47 ng/L. In the second approach the LOD was similar 8.62 ng/L, which is below of allowable limits of AFM1 in milk and milk products. The aptasensors were validated in a spiked milk samples with good recovery better than 78%. Comparative analysis of the sensitivity of immuno- and aptasensors was also performed and showed comparable sensitivity. Full article

(This article belongs to the Special Issue Membrane-Based Biosensing)

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14 pages, 5502 KiB

Open AccessArticle

Electrochemical DNA Sensor Based on Carbon Black—Poly(Neutral Red) Composite for Detection of Oxidative DNA Damage

by Yurii Kuzin, Dominika Kappo, Anna Porfireva, Dmitry Shurpik, Ivan Stoikov, Gennady Evtugyn and Tibor Hianik

Sensors 2018, 18(10), 3489; https://doi.org/10.3390/s18103489 - 16 Oct 2018

Cited by 31 | Viewed by 3357

Abstract

Voltammetric DNA sensor has been proposed on the platform of glassy carbon electrode covered with carbon black with adsorbed pillar[5]arene molecules. Electropolymerization of Neutral Red performed in the presence of native or oxidatively damaged DNA resulted in formation of hybrid material which activity depended on the DNA conditions. The assembling of the surface layer was confirmed by scanning electron microscopy and electrochemical impedance spectroscopy. The influence of DNA and pillar[5]arene on redox activity of polymeric dye was investigated and a significant increase of the peak currents was found for DNA damaged by reactive oxygen species generated by Cu²⁺/H₂O₂ mixture. Pillar[5]arene improves the electron exchange conditions and increases the response and its reproducibility. The applicability of the DNA sensor developed was shown on the example of ascorbic acid as antioxidant. It decreases the current in the concentration range from 1.0 μM to 1.0 mM. The possibility to detect antioxidant activity was qualitatively confirmed by testing tera infusion. The DNA sensor developed can find application in testing of carcinogenic species and searching for new antitumor drugs. Full article

(This article belongs to the Special Issue Membrane-Based Biosensing)

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19 pages, 9633 KiB

Open AccessArticle

Construction of Multiple Switchable Sensors and Logic Gates Based on Carboxylated Multi-Walled Carbon Nanotubes/Poly(N,N-Diethylacrylamide)

by Xuemei Wu, Xiaoqing Bai, Yang Ma, Jie Wei, Juan Peng, Keren Shi and Huiqin Yao

Sensors 2018, 18(10), 3358; https://doi.org/10.3390/s18103358 - 08 Oct 2018

Cited by 9 | Viewed by 3013

Abstract

In this work, binary hydrogel films based on carboxylated multi-walled carbon nanotubes/poly(N,N-diethylacrylamide) (c-MWCNTs/PDEA) were successfully polymerized and assembled on a glassy carbon (GC) electrode surface. The electroactive drug probes matrine and sophoridine in solution showed reversible thermal-, salt-, methanol- and pH-responsive switchable cyclic voltammetric (CV) behaviors at the film electrodes. The control experiments showed that the pH-responsive property of the system could be ascribed to the drug components of the solutions, whereas the thermal-, salt- and methanol-sensitive behaviors were attributed to the PDEA constituent of the films. The CV signals particularly, of matrine and sophoridine were significantly amplified by the electrocatalysis of c-MWCNTs in the films at 1.02 V and 0.91 V, respectively. Moreover, the addition of esterase, urease, ethyl butyrate, and urea to the solution also changed the pH of the system, and produced similar CV peaks as with dilution by HCl or NaOH. Based on these experiments, a 6-input/5-output logic gate system and 2-to-1 encoder were successfully constructed. The present system may lead to the development of novel types of molecular computing systems. Full article

(This article belongs to the Special Issue Membrane-Based Biosensing)

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29 pages, 2505 KiB

Open AccessArticle

The Role of Proton Transport in Gating Current in a Voltage Gated Ion Channel, as Shown by Quantum Calculations

by Alisher M. Kariev and Michael E. Green

Sensors 2018, 18(9), 3143; https://doi.org/10.3390/s18093143 - 18 Sep 2018

Cited by 6 | Viewed by 3841

Abstract

Over two-thirds of a century ago, Hodgkin and Huxley proposed the existence of voltage gated ion channels (VGICs) to carry Na⁺ and K⁺ ions across the cell membrane to create the nerve impulse, in response to depolarization of the membrane. The channels have multiple physiological roles, and play a central role in a wide variety of diseases when they malfunction. The first channel structure was found by MacKinnon and coworkers in 1998. Subsequently, the structure of a number of VGICs was determined in the open (ion conducting) state. This type of channel consists of four voltage sensing domains (VSDs), each formed from four transmembrane (TM) segments, plus a pore domain through which ions move. Understanding the gating mechanism (how the channel opens and closes) requires structures. One TM segment (S4) has an arginine in every third position, with one such segment per domain. It is usually assumed that these arginines are all ionized, and in the resting state are held toward the intracellular side of the membrane by voltage across the membrane. They are assumed to move outward (extracellular direction) when released by depolarization of this voltage, producing a capacitive gating current and opening the channel. We suggest alternate interpretations of the evidence that led to these models. Measured gating current is the total charge displacement of all atoms in the VSD; we propose that the prime, but not sole, contributor is proton motion, not displacement of the charges on the arginines of S4. It is known that the VSD can conduct protons. Quantum calculations on the K_v1.2 potassium channel VSD show how; the key is the amphoteric nature of the arginine side chain, which allows it to transfer a proton. This appears to be the first time the arginine side chain has had its amphoteric character considered. We have calculated one such proton transfer in detail: this proton starts from a tyrosine that can ionize, transferring to the NE of the third arginine on S4; that arginine’s NH then transfers a proton to a glutamate. The backbone remains static. A mutation predicted to affect the proton transfer has been qualitatively confirmed experimentally, from the change in the gating current-voltage curve. The total charge displacement in going from a normal closed potential of −70 mV across the membrane to 0 mV (open), is calculated to be approximately consistent with measured values, although the error limits on the calculation require caution in interpretation. Full article

(This article belongs to the Special Issue Membrane-Based Biosensing)

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11 pages, 5284 KiB

Open AccessArticle

Amperometric Detection of Conformational Change of Proteins Using Immobilized-Liposome Sensor System

by Hyunjong Yu, Young Hwan Son, Hak-Jin Kim, Keesung Kim, Pahn-Shick Chang and Ho-Sup Jung

Sensors 2018, 18(1), 136; https://doi.org/10.3390/s18010136 - 05 Jan 2018

Cited by 3 | Viewed by 4116

Abstract

An immobilized liposome electrode (ILE)-based sensor was developed to quantify conformational changes of the proteins under various stress conditions. The ILE surface was characterized by using a tapping-mode atomic force microscopy (TM-AFM) to confirm surface immobilization of liposome. The uniform layer of liposome was formed on the electrode. The current deviations generated based on the status of the proteins under different stress were then measured. Bovine carbonic anhydrase (CAB) and lysozyme were tested with three different conditions: native, reduced and partially denatured. For both proteins, a linear dynamic range formed between denatured concentrations and output electric current signals was able to quantify conformational changes of the proteins. The pattern recognition (PARC) technique was integrated with ILE-based sensor to perform data analysis and provided an effective method to improve the prediction of protein structural changes. The ILE-based stress sensor showed potential of leveraging the amperometric technique to manifest activity of proteins based on various external conditions. Full article

(This article belongs to the Special Issue Membrane-Based Biosensing)

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