scholarly journals Impedimetric and Plasmonic Sensing of Collagen I Using a Half-Antibody-Supported, Au-Modified, Self-Assembled Monolayer System

Biosensors ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 227
Author(s):  
Marcin Gwiazda ◽  
Sheetal K. Bhardwaj ◽  
Ewa Kijeńska-Gawrońska ◽  
Wojciech Swieszkowski ◽  
Unni Sivasankaran ◽  
...  
Keyword(s):  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
High Sensitivity ◽  
Covalent Immobilization ◽  
Electrochemical Immunosensor ◽  
Collagen I ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type I ◽  
Self Assembled Monolayer ◽  
Self Assembled

This research presents an electrochemical immunosensor for collagen I detection using a self-assembled monolayer (SAM) of gold nanoparticles (AuNPs) and covalently immobilized half-reduced monoclonal antibody as a receptor; this allowed for the validation of the collagen I concentration through two different independent methods: electrochemically by Electrochemical Impedance Spectroscopy (EIS), and optically by Surface Plasmon Resonance (SPR). The high unique advantage of the proposed sensor is based on the performance of the stable covalent immobilization of the AuNPs and enzymatically reduced half-IgG collagen I antibodies, which ensured their appropriate orientation onto the sensor’s surface, good stability, and sensitivity properties. The detection of collagen type I was performed in a concentration range from 1 to 5 pg/mL. Moreover, SPR was utilized to confirm the immobilization of the monoclonal half-antibodies and sensing of collagen I versus time. Furthermore, EIS experiments revealed a limit of detection (LOD) of 0.38 pg/mL. The selectivity of the performed immunosensor was confirmed by negligible responses for BSA. The performed approach of the immunosensor is a novel, innovative attempt that enables the detection of collagen I with very high sensitivity in the range of pg/mL, which is significantly lower than the commonly used enzyme-linked immunosorbent assay (ELISA).

scholarly journals Label-Free Electrochemical Biosensor Based on Au@MoS₂–PANI for Escherichia coli Detection

Chemosensors ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 49
Author(s):  
Pushap Raj ◽  
Man Hwan Oh ◽  
Kyudong Han ◽  
Tae Yoon Lee
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Limit Of Detection ◽  
Bacterial Pathogens ◽  
Cost Effective ◽  
Covalent Immobilization ◽  
Label Free ◽  
Self Assembled Monolayer ◽  
Detection Range ◽  
Linear Detection

Bacterial infections have become a significant challenge in terms of public health, the food industry, and the environment. Therefore, it is necessary to address these challenges by developing a rapid, cost-effective, and easy-to-use biosensor for early diagnosis of bacterial pathogens. Herein, we developed a simple, label-free, and highly sensitive immunosensor based on electrochemical detection using the Au@MoS₂–PANI nanocomposite. The conductivity of the glassy carbon electrode is greatly enhanced using the Au@MoS₂–PANI nanocomposite and a self-assembled monolayer of mercaptopropionic acid on the gold nanoparticle surface was employed for the covalent immobilization of antibodies to minimize the nonspecific adsorption of bacterial pathogens on the electrode surface. The biosensor established a high selectivity and sensitivity with a low limit of detection of 10 CFU/mL, and detected Escherichia coli within 30 min. Moreover, the developed biosensor demonstrated a good linear detection range, practical utility in urine samples, and electrode regenerative studies.

scholarly journals Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin

Toxins ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 458 ◽  
Author(s):  
Hisaya Ono ◽  
Nobuaki Hachiya ◽  
Yasunori Suzuki ◽  
Ikunori Naito ◽  
Shouhei Hirose ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Expression System ◽  
Sandwich Elisa ◽  
Food Poisoning ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
E Coli ◽  
Detection Systems ◽  
C Terminus ◽  
Culture Supernatants

Staphylococcal enterotoxins (SEs) are the cause of staphylococcal food poisoning (SFP) outbreaks. Recently, many new types of SEs and SE-like toxins have been reported, but it has not been proved whether these new toxins cause food poisoning. To develop an immunoassay for detection of SE-like J (SElJ), a non-characterized toxin in SFP, a mutant SElJ with C-terminus deletion (SElJ∆C) was expressed and purified in an E. coli expression system. Anti-SElJ antibody was produced in rabbits immunized with the SElJ∆C. Western blotting and sandwich enzyme-linked immunosorbent assay (ELISA) detection systems were established and showed that the antibody specifically recognizes SElJ without cross reaction to other SEs tested. The limit of detection for the sandwich ELISA was 0.078 ng/mL, showing high sensitivity. SElJ production in S. aureus was detected by using the sandwich ELISA and showed that selj-horboring isolates produced a large amount of SElJ in the culture supernatants, especially in that of the strain isolated from a food poisoning outbreak in Japan. These results demonstrate that the immunoassay for detection of SElJ is specific and sensitive and is useful for determining the native SElJ production in S. aureus isolated from food poisoning cases.

scholarly journals Dramatically Enhancing the Sensitivity of Immunoassay for Ochratoxin A Detection by Cascade-Amplifying Enzyme Loading

Toxins ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 781
Author(s):  
Zhuolin Song ◽  
Lin Feng ◽  
Yuankui Leng ◽  
Mingzhu Huang ◽  
Hao Fang ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Limit Of Detection ◽  
High Sensitivity ◽  
Cross Reaction ◽  
Molar Ratio ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Enzyme Loading ◽  
M13 Bacteriophage ◽  
Mimic Peptide

Enzyme-linked immunosorbent assay (ELISA) is widely used in the routine screening of mycotoxin contamination in various agricultural and food products. Herein, a cascade-amplifying system was introduced to dramatically promote the sensitivity of an immunoassay for ochratoxin A (OTA) detection. Specifically, a biotinylated M13 bacteriophage was introduced as a biofunctional competing antigen, in which a seven-peptide OTA mimotope fused on the p3 protein of M13 was used to specifically recognize an anti-OTA monoclonal antibody, and the biotin molecules modified on capsid p8 proteins were used in loading numerous streptavidin-labeled polymeric horseradish peroxidases (HRPs). Owing to the abundance of biotinylated p8 proteins in M13 and the high molar ratio between HRP and streptavidin in streptavidin-polyHRP, the loading amount of HRP enzymes on the M13 bacteriophage were greatly boosted. Hence, the proposed method exhibited high sensitivity, with a limit of detection of 2.0 pg/mL for OTA detection, which was 250-fold lower than that of conventional ELISA. In addition, the proposed method showed a slight cross-reaction of 2.3% to OTB, a negligible cross-reaction for other common mycotoxins, and an acceptable accuracy for OTA quantitative detection in real corn samples. The practicability of the method was further confirmed with a traditional HRP-based ELISA method. In conclusion, the biotinylated bacteriophage and polyHRP structure showed potential as a cascade-amplifying enzyme loading system for ultra-trace OTA detemination, and its application can be extended to the detection of other analytes by altering specific mimic peptide sequences.

Comparison of Proliferation and Growth of Human Keratinocytes on Plasma CO-Polymers of Acrylic acid/1,7- Octadiene and Self Assembled Monolayers

1998 ◽  
Vol 544 ◽  
Author(s):  
D. B. Haddow ◽  
R. M. France ◽  
R. D. Short ◽  
S. Macneil ◽  
R. A. Dawson
Keyword(s):  
Cell Proliferation ◽  
Tissue Culture ◽  
Carboxylic Acid ◽  
Cell Attachment ◽  
Human Keratinocytes ◽  
Collagen I ◽  
Self Assembled Monolayers ◽  
Self Assembled Monolayer ◽  
Assembled Monolayers ◽  
Self Assembled

AbstractHuman keratinocytes have been cultured on plasma co-polymers (PCPs), self assembled monolayers (SAMs), tissue culture poly(styrene) (TCPS) and collagen I. The degree of keratinocyte attachment was measured over 24 hours and cell proliferation and growth monitored over 7 days using optical microscopy and DNA concentrations. Cell attachment and proliferation and growth on the PCP surfaces were compared with 2 self assembled monolayer (SAM) systems. PCP surfaces containing carboxylic acid functionalities promoted keratinocyte attachment, with optimum attachment levels seen on surfaces containing less than 5% acid groups. The level of attachment on these surfaces was comparable to that seen on collagen I, a preferred substratum for the culturing of keratinocytes. After several days in culture the cells were well attached and proliferative. Keratinocytes attached well to acidterminated SAMs but attached poorly to methyl-terminated SAMs.

scholarly journals Impedance Sensing Platform for Detection of the Food Pathogen Listeria monocytogenes

Electronics ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 347 ◽  
Author(s):  
Maria Chiriacò ◽  
Ilaria Parlangeli ◽  
Fausto Sirsi ◽  
Palmiro Poltronieri ◽  
Elisabetta Primiceri
Keyword(s):  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Foodborne Pathogen ◽  
High Sensitivity ◽  
Primary Objective ◽  
Field Application ◽  
Impedance Sensing ◽  
Sensing Platform

A great improvement in food safety and quality controls worldwide has been achieved through the development of biosensing platforms. Foodborne pathogens continue to cause serious outbreaks, due to the ingestion of contaminated food. The development of new, sensitive, portable, high-throughput, and automated platforms is a primary objective to allow detection of pathogens and their toxins in foods. Listeria monocytogenes is one common foodborne pathogen. Major outbreaks of listeriosis have been caused by a variety of foods, including milk, soft cheeses, meat, fermented sausages, poultry, seafood and vegetable products. Due to its high sensitivity and easy setup, electrochemical impedance spectroscopy (EIS) has been extensively applied for biosensor fabrication and in particular in the field of microbiology as a mean to detect and quantify foodborne bacteria. Here we describe a miniaturized, portable EIS platform consisting of a microfluidic device with EIS sensors for the detection of L. monocytogenes in milk samples, connected to a portable impedance analyzer for on-field application in clinical and food diagnostics, but also for biosecurity purposes. To achieve this goal microelectrodes were functionalized with antibodies specific for L. monocytogenes. The binding and detection of L. monocytogenes was achieved in the range 2.2 × 103 cfu/mL to 1 × 102 with a Limit of Detection (LoD) of 5.5 cfu/mL.

Characterisation of electrochemical immunosensor for detection of viable not-culturable forms of Legionellla pneumophila in water samples

2015 ◽  
Vol 69 (11) ◽  
Author(s):  
Dejla Sboui ◽  
Mina Souiri ◽  
Stephanie Reynaud ◽  
Sabine Palle ◽  
Manel Ben Ismail ◽  
...  
Keyword(s):  
Legionella Pneumophila ◽  
Indium Tin Oxide ◽  
Membrane Filtration ◽  
Laser Scanning ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Angle Measurement ◽  
Electrochemical Immunosensor ◽  
Laser Scanning Microscopy ◽  
Confocal Laser

AbstractLegionella pneumophila may cause a fatal pneumonia in humans known as Legionnaires’ disease (LD). The strategies of L. pneumophila to adapt to and resist stressful environmental conditions include the ability to enter into a VBNC (viable but not culturable) state. The detection of L. pneumophila in environmental samples benefits from the use of standardised methods: for detection and enumeration following membrane filtration (AFNOR T90-431, ISO 11731) and detection and quantification by polymerase chain reaction PCR (AFNOR T90-471, ISO 12869). Culture is hampered by its inability to detect VBNC forms and PCR is unable to discriminate between live and dead bacteria. The present immunosensor was obtained by the immobilisation of a monoclonal anti-L. pneumophila antibody (MAb) on an indium-tin oxide (ITO) electrode by the self-assembled monolayers (SAMs) method using an aminosilane. The immunosensor was characterised by wettability (contact angle measurement), atomic force microscopy (AFM), confocal laser scanning microscopy (CLSM), and electrochemical impedance spectroscopy (EIS). A limit of detection of 10 bacteria per mL was observed on artificial samples.

scholarly journals Electrochemistry Study of Permselectivity and Interfacial Electron Transfers of a Branch-Tailed Fluorosurfactant Self-Assembled Monolayer on Gold

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 2998 ◽  
Author(s):  
Shanshan Li ◽  
Qingying Luo ◽  
Zhiqing Zhang ◽  
Guanghui Shen ◽  
Hejun Wu ◽  
...  
Keyword(s):  
Self Assembly ◽  
Electrochemical Impedance ◽  
Click Reaction ◽  
Hydration Shell ◽  
Self Assembled Monolayer ◽  
Tunneling Mechanism ◽  
Assembly Technique ◽  
Hydration Resistance ◽  
Self Assembled ◽  
Electron Transfers

We investigated the permselectivity and interfacial electron transfers of an amphiphilic branch-tailed fluorosurfactant self-assembled monolayer (FS-SAM) on a gold electrode by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The FS-SAM was prepared by a self-assembly technique and a “click” reaction. The barrier property and interfacial electron transfers of the FS-SAM were also evaluated using various probes with different features. The FS-SAM allowed a higher degree of permeation by small hydrophilic (Cl− and F−) electrolyte ions than large hydrophobic (ClO4− and PF6−) ones. Meanwhile, the redox reaction of the Fe(CN)63− couple was nearly completely blocked by the FS-SAM, whereas the electron transfer of Ru(NH3)63+ was easier than that of Fe(CN)63−, which may be due to the underlying tunneling mechanism. For hydrophobic dopamine, the hydrophobic bonding between the FS-SAM exterior fluoroalkyl moieties and the hydrophobic probes, as well as the hydration resistance from the interior hydration shell around the oligo (ethylene glycol) moieties, hindered the transport of hydrophobic probes into the FS-SAM. These results may have profound implications for understanding the permselectivity and electron transfers of amphiphilic surfaces consisting of molecules containing aromatic groups and branch-tailed fluorosurfactants in their structures.

scholarly journals Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

Biosensors ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 3
Author(s):  
Tiziano Di Giulio ◽  
Elisabetta Mazzotta ◽  
Cosimino Malitesta
Keyword(s):  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Mammalian Species ◽  
Covalent Immobilization ◽  
Amino Groups ◽  
Molecularly Imprinted ◽  
Disease Marker ◽  
Sensing Material ◽  
Enzymatic Marker ◽  
L 62

Herein we report the electropolymerization of a scopoletin based molecularly imprinted polymer (MIP) for the detection of lysozyme (Lyz), an enzymatic marker of several diseases in mammalian species. Two different approaches have been used for the imprinting of lysozyme based, respectively, on the use of a monomer-template mixture and on the covalent immobilization of the enzyme prior to polymer synthesis. In the latter case, a multi-step protocol has been exploited with preliminary functionalization of gold electrode with amino groups, via 4-aminothiophenol, followed by reaction with glutaraldehyde, to provide a suitable linker for lysozyme. Each step of surface electrode modification has been followed by cyclic voltammetry and electrochemical impedance spectroscopy, which has been also employed to test the electrochemical responses of the developed MIP. The sensors show good selectivity to Lyz and detect the enzyme at concentrations up to 292 mg/L (20 μM), but with different performances, depending on the used imprinting approach. An imprinting factor equal to 7.1 and 2.5 and a limit of detection of 0.9 mg/L (62 nM) and 2.1 mg/L (141 nM) have been estimated for MIPs prepared with and without enzyme immobilization, respectively. Competitive rebinding experiment results show that this sensing material is selective for Lyz determination. Tests were performed using synthetic saliva to evaluate the potential application of the sensors in real matrices for clinical purposes.

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