One-Dimensional Diffraction Grating of Poly(Methacrylic Acid) Brushes For The Rapid Label-Free Detection of Yersinia Pestis With a Reflective Laser System

Author(s):  
Feng-Ping Lin ◽  
Hui-Ling Hsu ◽  
Hui-Chung Lin ◽  
Hsin-Hsien Huang ◽  
Chien-Hsing Lu ◽  
...  

Abstract Background: Because of the low sensitivity of commercial products, development of a facile method to rapidly identify plague on-site remains highly attractive. Line arrays of poly(methacrylic acid) (PMAA) brushes were grafted using a photoresist template to fabricate one-dimensional diffraction gratings (DGs). The as-prepared samples first bound protein G to immobilize and orient the tails of the antibody of Yersinia pestis (abY). A laser beam was employed to analyze the 2D and 3D reflective signals of DGs at an incident angle of 45°. The abY-tailed PMAA DG possessed an optical feature with a characteristic diffraction effect along the SII, in which the projection of the laser beam on the plane of the DG chip was parallel to the strips, and ST configurations, in which they were perpendicular. A fluidic diffraction chip based on the abY-tailed PMMA DG was fabricated to examine the ability to detect Yersinia pestis along the ST configuration. Results: Upon flowing through the chip, Yersinia pestis was attached to the abY-tailed PMMA DG, which changed the diffraction intensity. The degree of the diffraction intensity exhibited a linear response to Yersinia pestis at concentrations from 102 to 107 CFU mL−1, and the limit of detection was 75 CFU mL−1, 1000 times lower than a commercial product (Alexter Bio-Detect Test). The diffractive sensor could selectively detect Yersinia pestis in spiked serum samples, with excellent standard deviation and recovery. Conclusion: Our platform provides a simple, label-free method for on-site plague diagnosis to prevent the highly rapid transmission of plague.

Nanophotonics ◽  
2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Gerardo A Lopez-Muñoz ◽  
Juan M Fernández-Costa ◽  
Maria Alejandra Ortega ◽  
Jordina Balaguer-Trias ◽  
Eduard Martin-Lasierra ◽  
...  

Abstract The development of nanostructured plasmonic biosensors has been widely widespread in the last years, motivated by the potential benefits they can offer in integration, miniaturization, multiplexing opportunities, and enhanced performance label-free biodetection in a wide field of applications. Between them, engineering tissues represent a novel, challenging, and prolific application field for nanostructured plasmonic biosensors considering the previously described benefits and the low levels of secreted biomarkers (≈pM–nM) to detect. Here, we present an integrated plasmonic nanocrystals-based biosensor using high throughput nanostructured polycarbonate substrates. Metallic film thickness and incident angle of light for reflectance measurements were optimized to enhance the detection of antibody–antigen biorecognition events using numerical simulations. We achieved an enhancement in biodetection up to 3× as the incident angle of light decreases, which can be related to shorter evanescent decay lengths. We achieved a high reproducibility between channels with a coefficient of variation below 2% in bulk refractive index measurements, demonstrating a high potential for multiplexed sensing. Finally, biosensing potential was demonstrated by the direct and label-free detection of interleukin-6 biomarker in undiluted cell culture media supernatants from bioengineered 3D skeletal muscle tissues stimulated with different concentrations of endotoxins achieving a limit of detection (LOD) of ≈ 0.03 ng/mL (1.4 pM).


Biosensors ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 180
Author(s):  
Lucia Sarcina ◽  
Giuseppe Felice Mangiatordi ◽  
Fabrizio Torricelli ◽  
Paolo Bollella ◽  
Zahra Gounani ◽  
...  

The early detection of the human immunodeficiency virus (HIV) is of paramount importance to achieve efficient therapeutic treatment and limit the disease spreading. In this perspective, the assessment of biosensing assay for the HIV-1 p24 capsid protein plays a pivotal role in the timely and selective detection of HIV infections. In this study, multi-parameter-SPR has been used to develop a reliable and label-free detection method for HIV-1 p24 protein. Remarkably, both physical and chemical immobilization of mouse monoclonal antibodies against HIV-1 p24 on the SPR gold detecting surface have been characterized for the first time. The two immobilization techniques returned a capturing antibody surface coverage as high as (7.5 ± 0.3) × 1011 molecule/cm2 and (2.4 ± 0.6) × 1011 molecule/cm2, respectively. However, the covalent binding of the capturing antibodies through a mixed self-assembled monolayer (SAM) of alkanethiols led to a doubling of the p24 binding signal. Moreover, from the modeling of the dose-response curve, an equilibrium dissociation constant KD of 5.30 × 10−9 M was computed for the assay performed on the SAM modified surface compared to a much larger KD of 7.46 × 10−5 M extracted for the physisorbed antibodies. The chemically modified system was also characterized in terms of sensitivity and selectivity, reaching a limit of detection of (4.1 ± 0.5) nM and an unprecedented selectivity ratio of 0.02.


2020 ◽  
Vol 13 (05) ◽  
pp. 2041004 ◽  
Author(s):  
Yang Li ◽  
Yanxian Guo ◽  
Binggang Ye ◽  
Zhengfei Zhuang ◽  
Peilin Lan ◽  
...  

Two-dimensional (2D) nanomaterials have captured an increasing attention in biophotonics owing to their excellent optical features. Herein, 2D hafnium ditelluride (HfTe[Formula: see text], a new member of transition metal tellurides, is exploited to support gold nanoparticles fabricating HfTe2-Au nanocomposites. The nanohybrids can serve as novel 2D surface-enhanced Raman scattering (SERS) substrate for the label-free detection of analyte with high sensitivity and reproducibility. Chemical mechanism originated from HfTe2 nanosheets and the electromagnetic enhancement induced by the hot spots on the nanohybrids may largely contribute to the superior SERS effect of HfTe2-Au nanocomposites. Finally, HfTe2-Au nanocomposites are utilized for the label-free SERS analysis of foodborne pathogenic bacteria, which realize the rapid and ultrasensitive Raman test of Escherichia coli, Listeria monocytogenes, Staphylococcus aureus and Salmonella with the limit of detection of 10 CFU/mL and the maximum Raman enhancement factor up to [Formula: see text]. Combined with principal component analysis, HfTe2-Au-based SERS analysis also completes the bacterial classification without extra treatment.


Polymers ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 4270
Author(s):  
Jem-Kun Chen ◽  
Feng-Ping Lin ◽  
Chi-Jung Chang ◽  
Chien-Hsing Lu ◽  
Chih-Feng Huang

Gelatin was immobilized selectively on the amide groups-modified bottom of a trench array of a photoresist template with 2 μm resolution by the ethyl(dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide reaction. The gelatin-immobilized line array was brominated to generate a macroinitiator for atom transfer radical polymerization. Poly(methacrylic acid) (PMAA) brushes were grafted from the macroinitiator layer as line arrays of one-dimensional diffraction gratings (DGs) for various grafting polymerization times. A laser beam system was employed to analyze the optical feature with a characteristic diffraction effect of the PMAA DGs at a 45° incident angle along the transverse magnetic and transverse electric polarization. The growth of the PMAA brush lines increased both their heights and widths, leading to a change in the reflective diffraction intensity. The PMAA brushes under various grafting polymerization times were cleaved from the substrate by digestion of gelatin with trypsin, and their molecular weights were obtained by gel permeation chromatography. The change degree of the diffraction intensity varied linearly with the molecular weight of the PMAA brushes over a wide range, from 135 to 1475 kDa, with high correlation coefficients. Molecular weight determination of polymer brushes using the reflective diffraction intensity provides a simple method to monitor their growth in real time without polymer brush cleavage.


Dengue virus (DENV) is a reemerging mosquito-borne disease that is endemic in more than 125 countries, affecting 200 million people per year. Screening testing has been a good attempt to minimize the impact caused by high morbity and mortality rates of DENV. In this study, a simple and disposable label-free electrochemical immunosensor based on a carbon ink graphite screen-printed electrode (SPE) one-step fabricated was developed for detection of non-structural 1 protein (NS1). The SPE surface was modified by drop casting, depositing a colloidal suspension containing amine-functionalized gold nanoparticles (AuNP-NH2). AuNPs were synthetized by a photoinduced physical method, illuminating preformed gold seeds with a light-emitting diode (LED,) at blue region, by using the polyethyleneimine (NH2) as reductor and stabilizing agent. UV-VIS spectroscopy and Transmission Electron Microscopy (TEM) were used to characterize the amine AuNPs. Electrocatalytic activity of AuNPs allowed more sensitivity for a label-free detection of NS1 by square wave voltammetry (SWV), with linear response from 0.1 to 2 µg mL-1. It was found a good linearity (coefficient of correlation of 0.995 (p<0.01) and a limit of detection of 0.03 µg mL-1 NS1 for analytical responses. AuNP-NH2 synthesis provided an easy oriented immobilization of anti-NS1 antibodies by Fc portion, resulting in a simple fabrication immunosensor with relative high performance and feasibility for early diagnostic of DENV.


2012 ◽  
Vol 2012 ◽  
pp. 1-12 ◽  
Author(s):  
Y. H. Huang ◽  
H. P. Ho ◽  
S. Y. Wu ◽  
S. K. Kong

Under certain conditions, a surface plasmon wave along a metal-dielectric interface can be excited by an optical beam. The reflected optical beam will then undergo changes in both intensity and phase. As the level of intensity or phase change is quite sensitive to the coupling conditions such as the molecule concentration on the metal surface, this phenomenon has been utilized for label-free detection of biological species and characterization of molecular interactions during the last two decades. Currently, most of the commercial surface plasmon resonance (SPR) sensors rely on the detection of absorption dip in angular or wavelength spectrum. However, recent researches have shown that phase detection has the potential to achieve lower limit of detection (LoD) and higher throughput. This paper, thus, intends to review various schemes and configurations for SPR phase detection. The performance advantages and disadvantages of various schemes will be emphasized. It is hoped that this paper will provide some insights to researchers interested in SPR sensing and help them to develop SPR sensors with better sensitivity and higher throughput.


2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Médéric Loyez ◽  
Mathilde Wells ◽  
Stéphanie Hambÿe ◽  
François Hubinon ◽  
Bertrand Blankert ◽  
...  

Abstract Background Early malaria diagnosis and its profiling require the development of new sensing platforms enabling rapid and early analysis of parasites in blood or saliva, aside the widespread rapid diagnostic tests (RDTs). Methods This study shows the performance of a cost-effective optical fiber-based solution to target the presence of Plasmodium falciparum histidine-rich protein 2 (PfHRP2). Unclad multimode optical fiber probes are coated with a thin gold film to excite Surface Plasmon Resonance (SPR) yielding high sensitivity to bio-interactions between targets and bioreceptors grafted on the metal surface. Results Their performances are presented in laboratory conditions using PBS spiked with growing concentrations of purified target proteins and within in vitro cultures. Two probe configurations are studied through label-free detection and amplification using secondary antibodies to show the possibility to lower the intrisic limit of detection. Conclusions As malaria hits millions of people worldwide, the improvement and multiplexing of this optical fiber technique can be of great interest, especially for a future purpose of using multiple receptors on the fiber surface or several coated-nanoparticles as amplifiers.


2021 ◽  
Vol 188 (3) ◽  
Author(s):  
Sofia Arshavsky-Graham ◽  
Anton Enders ◽  
Shanny Ackerman ◽  
Janina Bahnemann ◽  
Ester Segal

AbstractMicrofluidic integration of biosensors enables improved biosensing performance and sophisticated lab-on-a-chip platform design for numerous applications. While soft lithography and polydimethylsiloxane (PDMS)-based microfluidics are still considered the gold standard, 3D-printing has emerged as a promising fabrication alternative for microfluidic systems. Herein, a 3D-printed polyacrylate-based microfluidic platform is integrated for the first time with a label-free porous silicon (PSi)–based optical aptasensor via a facile bonding method. The latter utilizes a UV-curable adhesive as an intermediate layer, while preserving the delicate nanostructure of the porous regions within the microchannels. As a proof-of-concept, a generic model aptasensor for label-free detection of his-tagged proteins is constructed, characterized, and compared to non-microfluidic and PDMS-based microfluidic setups. Detection of the target protein is carried out by real-time monitoring reflectivity changes of the PSi, induced by the target binding to the immobilized aptamers within the porous nanostructure. The microfluidic integrated aptasensor has been successfully used for detection of a model target protein, in the range 0.25 to 18 μM, with a good selectivity and an improved limit of detection, when compared to a non-microfluidic biosensing platform (0.04 μM vs. 2.7 μM, respectively). Furthermore, a superior performance of the 3D-printed microfluidic aptasensor is obtained, compared to a conventional PDMS-based microfluidic platform with similar dimensions. Graphical abstract


2015 ◽  
Vol 33 (16) ◽  
pp. 3385-3393 ◽  
Author(s):  
Alberto Sinibaldi ◽  
Norbert Danz ◽  
Aleksei Anopchenko ◽  
Peter Munzert ◽  
Stefan Schmieder ◽  
...  

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1017 ◽  
Author(s):  
Reem Khan ◽  
Sondes Ben Aissa ◽  
Tauqir Sherazi ◽  
Gaelle Catanante ◽  
Akhtar Hayat ◽  
...  

In the present work, an aptasensing platform was developed for the detection of a carcinogenic mycotoxin termed patulin (PAT) using a label-free approach. The detection was mainly based on a specific interaction of an aptamer immobilized on carbon-based electrode. A long linear spacer of carboxy-amine polyethylene glycol chain (PEG) was chemically grafted on screen-printed carbon electrodes (SPCEs) via diazonium salt in the aptasensor design. The NH2-modified aptamer was then attached covalently to carboxylic acid groups of previously immobilized bifunctional PEG to build a diblock macromolecule. The immobilized diblocked molecules resulted in the formation of long tunnels on a carbon interface, while the aptamer was assumed as the gate of these tunnels. Upon target analyte binding, the gates were assumed to be closed due to conformational changes in the structure of the aptamer, increasing the resistance to the charge transfer. This increase in resistance was measured by electrochemical impedance spectroscopy, the main analytical technique for the quantitative detection of PAT. Encouragingly, a good linear range between 1 and 25 ng was obtained. The limit of detection and limit of quantification was 2.8 ng L−1 and 4.0 ng L−1, respectively. Selectivity of the aptasensor was confirmed with mycotoxins commonly occurring in food. The developed apta-assay was also applied to a real sample, i.e., fresh apple juice spiked with PAT, and toxin recovery up to 99% was observed. The results obtained validated the suitability and selectivity of the developed apta-assay for the identification and quantification of PAT in real food samples.


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