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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry".

Deadline for manuscript submissions: closed (15 August 2023) | Viewed by 14154

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Dr. Giorgio Rispoli

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Department of Neuroscience and rehabilitation, University of Ferrara, Via Luigi Borsari 46, 44121, Ferrara, Italy
Interests: nanostructured chemioresistive sensors; sensory receptor; Ionic transporter; signal transduction
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Dr. Pierpaolo Greco

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Department of Neuroscience and rehabilitation, University of Ferrara, Via Luigi Borsari 46, 44121, Ferrara, Italy
Interests: organic field effect transistors; organic electronics; surface plasmon resonance; Lab-on-chip
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Dear Colleagues,

Biosensors applied for the investigation of biomolecular interactions are still at the initial stage of development, and their successful development requires in-depth studies of both reaction kinetics and mechanisms of transduction. The large majority of previous research focused on applications in the fields of cancer and genomic research for the detection of disease biomarkers and genetic mutations.

Among the analytical techniques that are suitable for the quantification of the thermodynamic constants of binding are: systems based on mechanical and viscoelastic properties (functionalized cantilevers), optical-based biosensors with different schemes of exciting radiation and recording (plasmonic, fluorescence-based, coupled evanescent field), electrochemical devices with a discrete number of electrical protocols in continuous and alternate currents, electronic transducers, such as silicon nanowire transistors, organic semiconductors or conductive polymer-based transistors.

Lipid membranes constitute one of the main supporting systems for bioreceptor proteins in living organisms. Most of the reactions taking place during the recognition events reflect the exquisite selectivity and sensibility of the bioreceptor for particular analytes, and the complexity of the multicomponent environment that modifies the affinity of the inserted protein. Biosensors with integrated functional membranes mimicking the performances of bioreceptors are still in their infancy, and thus require extensive investigation.

Dr. Giorgio Rispoli
Dr. Pierpaolo Greco
Guest Editors

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Keywords

SPR biosensors
electrochemical biosensors
cantilever based biosensors
organic electronic based biosensors
lipid membranes

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New Trends in Molecular Biosensors in International Journal of Molecular Sciences

Published Papers (7 papers)

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Research

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17 pages, 7121 KiB

Open AccessArticle

Interaction of Ions in Organic and Aqueous Media with an Ion-Pair Sensor Equipped with a BODIPY Reporter: An ON1-OFF-ON2-ON3 Fluorescent Assay

by Marta Zaleskaya-Hernik, Łukasz Dobrzycki and Jan Romański

Int. J. Mol. Sci. 2023, 24(10), 8536; https://doi.org/10.3390/ijms24108536 - 10 May 2023

Cited by 2 | Viewed by 1180

Abstract

Here, we present a ditopic ion-pair sensor, B1, containing the BODIPY reporter unit in its structure, which is shown to be able—thanks to the presence of two heterogeneous binding domains—to interact with anions in an enhanced manner in the presence of cations. This enables it to interact with salts even in 99% aqueous solutions, making B1 a good candidate in terms of visual salt detection in the aquatic environment. Receptor B1’s ability to extract and release salt was applied in the transport of potassium chloride through a bulk liquid membrane. Working with a concentration of B1 in the organic phase and with the presence of a specific salt in an aqueous solution, an inverted transport experiment was also demonstrated. By varying the type and the amount of the anions added to B1, we were able to develop diverse optical responses, including a unique four-step ON1-OFF-ON2-ON3 output. Full article

(This article belongs to the Special Issue Molecular Biosensor)

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15 pages, 4818 KiB

Open AccessArticle

Direct Electrochemistry of Glucose Dehydrogenase-Functionalized Polymers on a Modified Glassy Carbon Electrode and Its Molecular Recognition of Glucose

by Yang Sun, Weishi Xue, Jianfeng Zhao, Qianqian Bao, Kailiang Zhang, Yupeng Liu and Hua Li

Int. J. Mol. Sci. 2023, 24(7), 6152; https://doi.org/10.3390/ijms24076152 - 24 Mar 2023

Cited by 2 | Viewed by 1836

Abstract

A glucose biosensor was layer-by-layer assembled on a modified glassy carbon electrode (GCE) from a nanocomposite of NAD(P)⁺-dependent glucose dehydrogenase, aminated polyethylene glycol (mPEG), carboxylic acid-functionalized multi-wall carbon nanotubes (fMWCNTs), and ionic liquid (IL) composite functional polymers. The electrochemical electrode was denoted as NF/IL/GDH/mPEG-fMWCNTs/GCE. The composite polymer membranes were characterized by cyclic voltammetry, ultraviolet-visible spectrophotometry, electrochemical impedance spectroscopy, scanning electron microscopy, and transmission electron microscopy. The cyclic voltammogram of the modified electrode had a pair of well-defined quasi-reversible redox peaks with a formal potential of −61 mV (vs. Ag/AgCl) at a scan rate of 0.05 V s⁻¹. The heterogeneous electron transfer constant (k_s) of GDH on the composite functional polymer-modified GCE was 6.5 s⁻¹. The biosensor could sensitively recognize and detect glucose linearly from 0.8 to 100 µM with a detection limit down to 0.46 μM (S/N = 3) and a sensitivity of 29.1 nA μM⁻¹. The apparent Michaelis–Menten constant (

K_{m}^{app}

) of the modified electrode was 0.21 mM. The constructed electrochemical sensor was compared with the high-performance liquid chromatography method for the determination of glucose in commercially available glucose injections. The results demonstrated that the sensor was highly accurate and could be used for the rapid and quantitative determination of glucose concentration. Full article

(This article belongs to the Special Issue Molecular Biosensor)

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14 pages, 1802 KiB

Open AccessArticle

Detection of Breast Cancer-Specific Extracellular Vesicles with Fiber-Optic SPR Biosensor

by Yagmur Yildizhan, Kaat Driessens, Hong Shen Kevin Tsao, Robin Boiy, Debby Thomas, Nick Geukens, An Hendrix, Jeroen Lammertyn and Dragana Spasic

Int. J. Mol. Sci. 2023, 24(4), 3764; https://doi.org/10.3390/ijms24043764 - 13 Feb 2023

Cited by 4 | Viewed by 2482

Abstract

Extracellular vesicles (EVs) have attracted great attention as potential biomarkers for cancer diagnostics. Although several technologies have been developed for EV detection, many of them are still not applicable to clinical settings as they rely on complex EV isolation processes, while lacking sensitivity, specificity or standardization. To solve this problem, we have developed a sensitive breast cancer-specific EV detection bioassay directly in blood plasma using a fiber-optic surface plasmon resonance (FO-SPR) biosensor, previously calibrated with recombinant EVs. First, we established a sandwich bioassay to detect SK-BR-3 EVs by functionalizing the FO-SPR probes with anti-HER2 antibodies. A calibration curve was built using an anti-HER2/^Banti-CD9 combination, resulting in an LOD of 2.1 × 10⁷ particles/mL in buffer and 7 × 10⁸ particles/mL in blood plasma. Next, we investigated the potential of the bioassay to detect MCF7 EVs in blood plasma using an anti-EpCAM/^Banti-mix combination, obtaining an LOD of 1.1 × 10 ⁸ particles/mL. Finally, the specificity of the bioassay was proven by the absence of signal when testing plasma samples from 10 healthy people unknown to be diagnosed with breast cancer. The remarkable sensitivity and specificity of the developed sandwich bioassay together with the advantages of the standardized FO-SPR biosensor highlight outstanding potential for the future of EV analysis. Full article

(This article belongs to the Special Issue Molecular Biosensor)

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12 pages, 2609 KiB

Open AccessArticle

Modular Site-Specific Conjugation of Nanobodies Using Two Co-Associating Tags

by Eric Moeglin, Lina Barret, Bruno Chatton and Mariel Donzeau

Int. J. Mol. Sci. 2022, 23(22), 14405; https://doi.org/10.3390/ijms232214405 - 19 Nov 2022

Cited by 3 | Viewed by 1667

Abstract

The homogeneous labeling of antibodies and their fragments is a critical step for the generation of robust probes used in immuno-detection applications. To date, numerous chemical, genetic and peptide-based site-specific coupling methods have been developed. Among these methods, co-assembling peptide-tags is one of the most straightforward and versatile solutions. Here, we describe site-specific labeling of nanobodies through the use of two co-associating peptides tags, E3 and K3, originating from the tetramerization domain of p53. These E3 and K3-tags provide a simple and robust method for associating stoichiometric amount of V_HH and fluorescent probes, either fluorescent proteins or fluorochromes, at specific positions. As a proof of concept, a nanobody targeting the human epidermal growth factor receptor 2 (HER2), the nano-HER2 was genetically fused to the E3 and associated with different fluorescent K3-derivates. Entities were produced separately in Escherichia coli in soluble forms at high yields and co-assembled in vitro. These molecular probes present high binding specificity on HER2-overexpressing cells in flow-cytometry with relative binding constants in the low nanomolar range and are stable enough to stain HER2-receptor on living cells followed detection using fluorescent confocal microscopy. Altogether, our results demonstrate that the non-covalent conjugation method using these two co-associating peptides can be easily implemented for the modular engineering of molecular probes for cell immuno-staining. Full article

(This article belongs to the Special Issue Molecular Biosensor)

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13 pages, 3179 KiB

Open AccessArticle

Electrochemical Biosensor Designed to Distinguish Tetracyclines Derivatives by ssDNA Aptamer Labelled with Ferrocene

by Kamila Malecka-Baturo, Apostolos Zaganiaris, Iwona Grabowska and Katarzyna Kurzątkowska-Adaszyńska

Int. J. Mol. Sci. 2022, 23(22), 13785; https://doi.org/10.3390/ijms232213785 - 09 Nov 2022

Cited by 2 | Viewed by 1550

Abstract

Controlling food safety and preventing the growing spread of antibiotics into food products have been challenging problems for the protection of human health. Hence, the development of easy-to-use, fast, and sensitive analytical methods for the detection of antibiotics in food products has become one of the priorities in the food industry. In this paper, an electrochemical platform based on the ssDNA aptamer for the selective detection of tetracycline has been proposed. The aptasensor is based on a thiolated aptamer, labelled with ferrocene, which has been covalently co-immobilized onto a gold electrode surface with 6-mercaptohexan-1-ol. The changes in the redox activity of ferrocene observed on the aptamer–antibiotics interactions have been the basis of analytical signal generation registered by square-wave voltammetry. Furthermore, the detection of tetracycline-spiked cow milk samples has been successfully demonstrated. The limits of detection (LODs) have been obtained of 0.16 nM and 0.20 nM in the buffer and spiked cow milk, respectively, which exceed the maximum residue level (225 nM) more than 1000 times. The proposed aptasensor offers high selectivity for tetracycline against other structurally related tetracycline derivatives. The developed biosensor characterized by simplicity, a low detection limit, and high reliability shows practical potential for the detection of tetracycline in animal-origin milk. Full article

(This article belongs to the Special Issue Molecular Biosensor)

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10 pages, 2337 KiB

Open AccessCommunication

Rapid Colorimetric Detection of Wound Infection with a Fluidic Paper Device

by Javier Hoyo, Arnau Bassegoda, Guillem Ferreres, Dolores Hinojosa-Caballero, Manuel Gutiérrez-Capitán, Antoni Baldi, César Fernández-Sánchez and Tzanko Tzanov

Int. J. Mol. Sci. 2022, 23(16), 9129; https://doi.org/10.3390/ijms23169129 - 15 Aug 2022

Cited by 3 | Viewed by 1807

Abstract

Current procedures for the assessment of chronic wound infection are time-consuming and require complex instruments and trained personnel. The incidence of chronic wounds worldwide, and the associated economic burden, urge for simple and cheap point-of-care testing (PoCT) devices for fast on-site diagnosis to enable appropriate early treatment. The enzyme myeloperoxidase (MPO), whose activity in infected wounds is about ten times higher than in non-infected wounds, appears to be a suitable biomarker for wound infection diagnosis. Herein, we develop a single-component foldable paper-based device for the detection of MPO in wound fluids. The analyte detection is achieved in two steps: (i) selective immunocapture of MPO, and (ii) reaction of a specific dye with the captured MPO, yielding a purple color with increasing intensity as a function of the MPO activity in infected wounds in the range of 20–85 U/mL. Ex vivo experiments with wound fluids validated the analytic efficiency of the paper-based device, and the results strongly correlate with a spectrophotometric assay. Full article

(This article belongs to the Special Issue Molecular Biosensor)

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Review

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20 pages, 1580 KiB

Open AccessReview

Recent Developments in Electrochemical-Impedimetric Biosensors for Virus Detection

by Zala Štukovnik and Urban Bren

Int. J. Mol. Sci. 2022, 23(24), 15922; https://doi.org/10.3390/ijms232415922 - 14 Dec 2022

Cited by 15 | Viewed by 3016

Abstract

Viruses, including influenza viruses, MERS-CoV (Middle East respiratory syndrome coronavirus), SARS-CoV (severe acute respiratory syndrome coronavirus), HAV (Hepatitis A virus), HBV (Hepatitis B virus), HCV (Hepatitis C virus), HIV (human immunodeficiency virus), EBOV (Ebola virus), ZIKV (Zika virus), and most recently SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), are responsible for many diseases that result in hundreds of thousands of deaths yearly. The ongoing outbreak of the COVID-19 disease has raised a global concern and intensified research on the detection of viruses and virus-related diseases. Novel methods for the sensitive, rapid, and on-site detection of pathogens, such as the recent SARS-CoV-2, are critical for diagnosing and treating infectious diseases before they spread and affect human health worldwide. In this sense, electrochemical impedimetric biosensors could be applied for virus detection on a large scale. This review focuses on the recent developments in electrochemical-impedimetric biosensors for the detection of viruses. Full article

(This article belongs to the Special Issue Molecular Biosensor)

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