Rapid, targeted and culture-free viral infectivity assay in drop-based microfluidics

Lab on a Chip ◽  
2015 ◽  
Vol 15 (19) ◽  
pp. 3934-3940 ◽  
Author(s):  
Ye Tao ◽  
Assaf Rotem ◽  
Huidan Zhang ◽  
Connie B. Chang ◽  
Anindita Basu ◽  
...  
Keyword(s):  
High Throughput ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Cost Effective ◽  
Serial Dilution ◽  
Viral Infectivity ◽  
Large Dynamic Range ◽  
Infectivity Assay ◽  
Dilution Experiments

We developed a rapid, targeted and culture-free infectivity assay using high-throughput drop-based microfluidics. The high sensitivity and large dynamic range of our cost effective assay alleviates the need for serial dilution experiments.

scholarly journals Rapid and Sensitive Quantification of Intracellular Glycyl-Sarcosine for Semi-High-Throughput Screening for Inhibitors of PEPT-1

Pharmaceutics ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1019
Author(s):  
Teresa von Linde ◽  
Gzona Bajraktari-Sylejmani ◽  
Walter E. Haefeli ◽  
Jürgen Burhenne ◽  
Johanna Weiss ◽  
...  
Keyword(s):  
Sample Preparation ◽  
High Throughput ◽  
High Throughput Screening ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Peptide Bond ◽  
Peptide Transporter ◽  
Systemic Absorption ◽  
Screening Assay ◽  
Limit Of Quantification

The peptide transporter PEPT-1 (SLC15A1) plays a major role in nutritional supply with amino acids by mediating the intestinal influx of dipeptides and tripeptides generated during food digestion. Its role in the uptake of small bioactive peptides and various therapeutics makes it an important target for the investigation of the systemic absorption of small peptide-like active compounds and prodrug strategies of poorly absorbed therapeutics. The dipeptide glycyl-sarcosine (Gly-Sar), which comprises an N-methylated peptide bond that increases stability against enzymatic degradation, is widely utilized for studying PEPT-1-mediated transport. To support experiments on PEPT-1 inhibitor screening to identify potential substrates, we developed a highly sensitive Gly-Sar quantification assay for Caco-2 cell lysates with a dynamic range of 0.1 to 1000 ng/mL (lower limit of quantification 0.68 nM) in 50 µL of cell lysate. The assay was validated following the applicable recommendations for bioanalytic method validation of the FDA and EMA. Sample preparation and quantification were established in 96-well cell culture plates that were also used for the cellular uptake studies, resulting in a rapid and robust screening assay for PEPT-1 inhibitors. This sample preparation principle, combined with the high sensitivity of the UPLC-MS/MS quantification, is suitable for screening assays for PEPT-1 inhibitors and substrates in high-throughput formats and holds the potential for automation. Applicability was demonstrated by IC50 determinations of the known PEPT-1 inhibitor losartan, the known substrates glycyl-proline (Gly-Pro), and valaciclovir, the prodrug of aciclovir, which itself is no substrate of PEPT-1 and consequently showed no inhibition in our assay.

The Design and Simulation of a New Z-Axis Resonant Micro-Accelerometer Based on Electrostatic Stiffness

2013 ◽  
Vol 744 ◽  
pp. 478-483
Author(s):  
Bo Yang ◽  
Bo Dai ◽  
Hui Zhao
Keyword(s):  
Dynamic Range ◽  
System Simulation ◽  
High Sensitivity ◽  
Scale Factor ◽  
Effective Frequency ◽  
Large Dynamic Range ◽  
Digital Output ◽  
Structure Simulation ◽  
Simulation Results ◽  
Micro Accelerometer

Resonant micro-accelerometers have good properties such as the large dynamic range, the high sensitivity, the strong anti-interference ability as well as the direct digital output. A new z-axis resonant micro-accelerometer based on electrostatic stiffness is researched. The new z-axis resonant micro-accelerometer consists of a torsional accelerometer and two plane resonators. The sensing movement of the accelerometer is decoupled with oscillation of the plane resonators by electrostatic stiffness, which will benefit to improve the performance of the new z-axis resonant micro-accelerometer. The new structure is designed. The sensitive theory of the acceleration is investigated and the equation of scale factor is deduced under ideal conditions. The simulation is implemented to verify the basic principle by the Ansys and Matlab. The structure simulation results prove that the effective frequency of the torsional accelerometer and the resonator are 0.66kHz and 13.3kHz separately. And the interference modes are isolated with the effective mode apparently. The system simulation results indicate that the scale factor is 37Hz/g and the system has excellent capabilities in locking and tracking natural frequency of resonators, which proves that the basic theory is feasible.

scholarly journals Theoretical Analysis of a Microring Resonator Array with High Sensitivity and Large Dynamic Range Based on a Multi-Scale Technique

Sensors ◽  
2018 ◽  
Vol 18 (7) ◽  
pp. 1987
Author(s):  
Wenqin Mo ◽  
Huiyun Liu ◽  
Fang Jin ◽  
Junlei Song ◽  
Kaifeng Dong
Keyword(s):  
Theoretical Analysis ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Microring Resonator ◽  
Large Dynamic Range ◽  
Multi Scale

scholarly journals Development of a high-throughput homogeneous AlphaLISA drug screening assay for the detection of SARS-CoV-2 Nucleocapsid

2020 ◽  
Author(s):  
Kirill Gorshkov ◽  
Catherine Z. Chen ◽  
Juan Carlos de la Torre ◽  
Luis Martinez-Sobrido ◽  
Thomas Moran ◽  
...  
Keyword(s):  
Tissue Culture ◽  
High Throughput ◽  
High Throughput Screening ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Research Field ◽  
Screening Assay ◽  
Detection Technology ◽  
Drug Screening Assay ◽  
Culture Supernatants

AbstractThe coronavirus disease 2019 (COVID-19) pandemic caused by Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) is in urgent need of therapeutic options. High-throughput screening (HTS) offers the research field an opportunity to rapidly identify such compounds. In this work, we have developed a homogeneous cell-based HTS system using AlphaLISA detection technology for the SARS-CoV-2 nucleocapsid protein (NP). Our assay measures both recombinant NP and endogenous NP from viral lysates and tissue culture supernatants (TCS) in a sandwich-based format using two monoclonal antibodies against the NP analyte. Viral NP was detected and quantified in both tissue culture supernatants and cell lysates, with large differences observed between 24 hours and 48 hours of infection. We simulated the viral infection by spiking in recombinant NP into 384-well plates with live Vero-E6 cells and were able to detect the NP with high sensitivity and a large dynamic range. Anti-viral agents that inhibit either viral cell entry or replication will decrease the AlphaLISA NP signal. Thus, this assay can be used for high-throughput screening of small molecules and biologics in the fight against the COVID-19 pandemic.

A novel approach to design electrochemical aptamer-based biosensor for ultrasensitive detecting of zearalenone as a prevalent estrogenic mycotoxin

2021 ◽  
Vol 28 ◽  
Author(s):  
Shokoufeh Hassani ◽  
Armin Salek Maghsoudi ◽  
Milad Rezaei Akmal ◽  
Shahram Shoeibi ◽  
Fatemeh Ghadipasha ◽  
...  
Keyword(s):  
Dynamic Range ◽  
Electrochemical Impedance ◽  
Fusarium Species ◽  
High Sensitivity ◽  
High Specificity ◽  
Cost Effective ◽  
Differential Pulse ◽  
Electrode Position ◽  
Novel Approach ◽  
Modified Surface

Background: Zearalenone is a well-known estrogenic mycotoxin produced by Fusarium species, a serious threat to the agricultural and food industries worldwide. Zearalenone, with its known metabolites, are biomarkers of exposure to certain fungi, primarily through food. It has considerable toxic effects on biological systems due to its carcinogenicity, mutagenicity, renal toxicity, teratogenicity, and immunotoxicity. Introduction: This study aims to design a simple, quick, precise, and cost-effective method on a biosensor platform to evaluate the low levels of this toxin in foodstuffs and agricultural products. Methods: An aptamer-based electrochemical biosensor was introduced that utilizes screen-printed gold electrodes instead of conventional electrodes. The electrode position process was employed to develop a gold nanoparticle-modified surface to enhance the electroactive surface area. Thiolated aptamers were immobilized on the surface of gold nanoparticles, and subsequently, the blocker and analyte were added to the modified surface. In the presence of a redox probe, electrochemical characterization of differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectroscopy were used to investigate the various stages of aptasensor fabrication. Results: The proposed aptasensor for zearalenone concentration had a wide linear dynamic range covering the 0.5 pg/mL to 100 ng/mL with a 0.14 pg/mL detection limit. Moreover, this aptasensor had high specificity so that a non-specific analyte cannot negatively affect the selectivity of the aptasensor. Conclusion: Overall, due to its simple design, high sensitivity, and fast performance, this aptasensor showed a high potential for assessing zearalenone in real samples, providing a clear perspective for designing a portable and cost-effective device.

scholarly journals High-sensitivity and large-dynamic-range refractive index sensors employing weak composite Fabry–Perot cavities

2017 ◽  
Vol 42 (16) ◽  
pp. 3145 ◽  
Author(s):  
Pengcheng Chen ◽  
Xuewen Shu ◽  
Haoran Cao ◽  
Kate Sugden
Keyword(s):  
Refractive Index ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Large Dynamic Range ◽  
Fabry Perot ◽  
Refractive Index Sensors
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