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High-throughput screening

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https://www.readbyqxmd.com/read/29786885/automated-buildup-of-biomimetic-films-in-cell-culture-microplates-for-high-throughput-screening-of-cellular-behaviors
#1
Paul Machillot, Catarina Quintal, Fabien Dalonneau, Loic Hermant, Pauline Monnot, Kelsey Matthews, Vincent Fitzpatrick, Jie Liu, Isabelle Pignot-Paintrand, Catherine Picart
An automatic method is established for layer-by-layer (LbL) assembly of biomimetic coatings in cell culture microplates using a commercial liquid-handling robot. Highly homogeneous thin films are formed at the bottom of each microwell. The LbL film-coated microplates are compatible with common cellular assays, using microplate readers and automated microscopes. Cellular adhesion is screened on crosslinked and peptide-functionalized LbL films and stem cell differentiation in response to increasing doses of bone morphogenetic proteins (2, 4, 7, 9)...
May 22, 2018: Advanced Materials
https://www.readbyqxmd.com/read/29786565/modelling-non-alcoholic-fatty-liver-disease-in-human-hepatocyte-like-cells
#2
Marcus J Lyall, Jessy Cartier, John P Thomson, Kate Cameron, Jose Meseguer-Ripolles, Eoghan O'Duibhir, Dagmara Szkolnicka, Baltasar Lucendo Villarin, Yu Wang, Giovanny Rodriguez Blanco, Warwick B Dunn, Richard R Meehan, David C Hay, Amanda J Drake
Non-alcoholic fatty liver disease (NAFLD) is the most common cause of liver disease in developed countries. An in vitro NAFLD model would permit mechanistic studies and enable high-throughput therapeutic screening. While hepatic cancer-derived cell lines are a convenient, renewable resource, their genomic, epigenomic and functional alterations mean their utility in NAFLD modelling is unclear. Additionally, the epigenetic mark 5-hydroxymethylcytosine (5hmC), a cell lineage identifier, is rapidly lost during cell culture, alongside expression of the Ten-eleven-translocation ( TET ) methylcytosine dioxygenase enzymes, restricting meaningful epigenetic analysis...
July 5, 2018: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/29786562/application-of-hepatocyte-like-cells-to-enhance-hepatic-safety-risk-assessment-in-drug-discovery
#3
REVIEW
Dominic P Williams
Hepatic stress and injury from drugs continues to be a major concern within the pharmaceutical industry, leading to preclinical and clinical attrition precautionary warnings and post-market withdrawal of drugs. There is a requirement for more predictive and mechanistically accurate models to aid risk assessment. Primary human hepatocytes, subject to isolation stress, cryopreservation, donor-to-donor variation and a relatively short period of functional capability in two-dimensional cultures, are not suitable for high-throughput screening procedures...
July 5, 2018: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/29785715/current-advances-in-screening-for-bioactive-components-from-medicinal-plants-by-affinity-ultrafiltration-mass-spectrometry
#4
Guilin Chen, Bill X Huang, Mingquan Guo
INTRODUCTION: Medicinal plants have played an important role in maintaining human health for thousands of years. However, the interactions between the active components in medicinal plants and some certain biological targets during a disease are still unclear in most cases. OBJECTIVE: To conduct the high-throughput screening for small active molecules that can interact with biological targets, which is of great theoretical significance and practical value. METHODOLOGY: The ultrafiltration mass spectrometry (UF-LC/MS) is a powerful bio-analytical method by combining affinity ultrafiltration and liquid chromatography-mass spectrometry (LC/MS), which could rapidly screen and identify small active molecules that bind to biological targets of interest at the same time...
May 21, 2018: Phytochemical Analysis: PCA
https://www.readbyqxmd.com/read/29784948/simultaneous-cell-disruption-and-semi-quantitative-activity-assays-for-high-throughput-screening-of-thermostable-l-asparaginases
#5
Xu Li, Xian Zhang, Shuqin Xu, Hengwei Zhang, Meijuan Xu, Taowei Yang, Li Wang, Haifeng Qian, Huiling Zhang, Haitian Fang, Tolbert Osire, Zhiming Rao, Shangtian Yang
L-asparaginase, which catalyses the hydrolysis of L-asparagine to L-aspartate, has attracted the attention of researchers due to its expanded applications in medicine and the food industry. In this study, a novel thermostable L-asparaginase from Pyrococcus yayanosii CH1 was cloned and over-expressed in Bacillus subtilis 168. To obtain thermostable L-asparaginase mutants with higher activity, a robust high-throughput screening process was developed specifically for thermophilic enzymes. In this process, cell disruption and enzyme activity assays are simultaneously performed in 96-deep well plates...
May 21, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29784937/an-oil-free-picodrop-bioassay-platform-for-synthetic-biology
#6
Christian A Siltanen, Russell H Cole, Sean Poust, Lawrence Chao, Jabus Tyerman, Benjamin Kaufmann-Malaga, Jeff Ubersax, Zev J Gartner, Adam R Abate
Droplet microfluidics enables massively-parallel analysis of single cells, biomolecules, and chemicals, making it valuable for high-throughput screens. However, many hydrophobic analytes are soluble in carrier oils, preventing their quantitative analysis with the method. We apply Printed Droplet Microfluidics to construct defined reactions with chemicals and cells incubated under air on an open array. The method interfaces with most bioanalytical tools and retains hydrophobic compounds in compartmentalized reactors, allowing their quantitation...
May 21, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29784819/xenoprotein-engineering-via-synthetic-libraries
#7
Zachary P Gates, Alexander A Vinogradov, Anthony J Quartararo, Anupam Bandyopadhyay, Zi-Ning Choo, Ethan D Evans, Kathryn H Halloran, Alexander J Mijalis, Surin K Mong, Mark D Simon, Eric A Standley, Evan D Styduhar, Sarah Z Tasker, Faycal Touti, Jessica M Weber, Jessica L Wilson, Timothy F Jamison, Bradley L Pentelute
Chemical methods have enabled the total synthesis of protein molecules of ever-increasing size and complexity. However, methods to engineer synthetic proteins comprising noncanonical amino acids have not kept pace, even though this capability would be a distinct advantage of the total synthesis approach to protein science. In this work, we report a platform for protein engineering based on the screening of synthetic one-bead one-compound protein libraries. Screening throughput approaching that of cell surface display was achieved by a combination of magnetic bead enrichment, flow cytometry analysis of on-bead screens, and high-throughput MS/MS-based sequencing of identified active compounds...
May 21, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29784672/genetic-screening-and-multipotency-in-rhesus-monkey-haploid-neural-progenitor-cells
#8
Haisong Wang, Wenhao Zhang, Jian Yu, Congyu Wu, Qian Gao, Xu Li, Yanni Li, Jinxin Zhang, Yaru Tian, Tao Tan, Weizhi Ji, Luyuan Li, Yang Yu, Ling Shuai
Haploid embryonic stem cells (haESCs) have been extensively applied in forward and reverse genetic screening. However, the mammalian haploid somatic cell line is difficult to achieve because of spontaneous diploidization in differentiation. As a non-human primate species, monkeys are widely used in basic and pre-clinical research in which haploid cells are restricted to ESCs. Here, we report that rhesus monkey haESCs in an optimized culture medium showed naïve-state pluripotency and stable haploidy. This model facilitated the derivation of haploid neural progenitor cells (haNPCs), which maintained haploidy and differentiation potential into neurons and glia for a long period in vitro High-throughput trapping mutations can be efficiently introduced into haNPCs via piggyBac transposons...
May 21, 2018: Development
https://www.readbyqxmd.com/read/29784363/a-colorimetric-immunoassay-based-on-glucose-oxidase-induced-aunp-aggregation-for-the-detection-of-fumonisin-b-1
#9
Xirui Chen, Yi Liang, Wenjing Zhang, Yuankui Leng, Yonghua Xiong
Naked-eye readout colorimetric signal-based assays are promising for high-throughput screening detection and point-of-care diagnostics in resource-constrained regions. Here, a novel direct competitive plasmonic enzyme-linked immunosorbent assay (dc-pELISA) based on gold nanoparticle (AuNP) aggregation with highly sensitive and robust naked-eye readout signal was developed and used to detect fumonisin B1 (FB1 ). AuNP aggregation was induced by a horseradish peroxidase (HRP)/hydrogen peroxide (H2 O2 )/tyramine (TYR) system, resulting in a dramatic color change from red to blue...
August 15, 2018: Talanta
https://www.readbyqxmd.com/read/29784362/programmable-electrochemical-flow-system-for-high-throughput-determination-of-total-antioxidant-capacity
#10
Kornautchaya Veenuttranon, Loc Thai Nguyen
The aim of this work was to develop a programmable flow system for rapid assessment of total antioxidant capacity (TAC). Novel features of the prototype include a single pressure-driven source, versatile manipulation of fluid flows, ability to adapt to different TAC assays, and compatibility with microfluidic design. Antioxidant activity was determined by electrochemical measurement of residual 2,2-diphenyl-2-picrylhydrayl hydrate (DPPH•) free radicals in the solution. The overall performance of the device was validated by the spectrophotometric method...
August 15, 2018: Talanta
https://www.readbyqxmd.com/read/29782834/high-throughput-dissection-of-aav-host-interactions-the-fast-and-the-curious
#11
REVIEW
Anne-Kathrin Herrmann, Dirk Grimm
Over fifty years after its initial description, Adeno-associated virus (AAV) remains a most exciting but also most elusive study object in basic or applied virology. On the one hand, its simple structure not only facilitates investigations into virus biology, but combined with the availability of numerous natural AAV variants with distinct infection efficiency and specificity also makes AAV a preferred substrate for engineering of gene delivery vectors. On the other hand, it is striking to witness a recent flurry of reports that highlight and partially close persistent gaps in our understanding of AAV virus and vector biology...
May 18, 2018: Journal of Molecular Biology
https://www.readbyqxmd.com/read/29782484/implications-for-human-odor-sensing-revealed-from-the-statistics-of-odorant-receptor-interactions
#12
Ji Hyun Bak, Seogjoo J Jang, Changbong Hyeon
Binding of odorants to olfactory receptors (ORs) elicits downstream chemical and neural signals, which are further processed to odor perception in the brain. Recently, Mainland and colleagues have measured more than 500 pairs of odorant-OR interaction by a high-throughput screening assay method, opening a new avenue to understanding the principles of human odor coding. Here, using a recently developed minimal model for OR activation kinetics, we characterize the statistics of OR activation by odorants in terms of three empirical parameters: the half-maximum effective concentration EC50, the efficacy, and the basal activity...
May 21, 2018: PLoS Computational Biology
https://www.readbyqxmd.com/read/29781496/a-universal-platform-for-selection-and-high-resolution-phenotypic-screening-of-bacterial-mutants-using-the-nanowell-slide
#13
H Antypas, M Veses-Garcia, E Weibull, H Andersson-Svahn, A Richter-Dahlfors
The Petri dish and microtiter plate are the golden standard for selection and screening of bacteria in microbiological research. To improve on the limited resolution and throughput of these methods, we developed a universal, user-friendly platform for selection and high-resolution phenotypic screening based on the nanowell slide. This miniaturized platform has an optimal ratio between throughput and assay complexity, holding 672 nanowells of 500 nl each. As monoclonality is essential in bacterial genetics, we used FACS to inoculate each nanowell with a single bacterium in 15 min...
May 21, 2018: Lab on a Chip
https://www.readbyqxmd.com/read/29781014/cat-elcca-catalyzing-drug-discovery-through-click-chemistry
#14
Amanda L Garner
Click chemistry has emerged as a powerful tool in our arsenal for unlocking new biology. This includes its utility in both chemical biology and drug discovery. An emerging application of click chemistry is in the development of biochemical assays for high-throughput screening to identify new chemical probes and drug leads. This Feature Article will discuss the advancements in click chemistry that were necessary for the development of a new class of biochemical assay, catalytic enzyme-linked click chemistry assay or cat-ELCCA...
May 21, 2018: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/29781002/high-throughput-screening-of-solid-state-catalysts-for-nerve-agent-degradation
#15
Joseph M Palomba, Cy V Credille, Mark Kalaj, Jared B DeCoste, Gregory W Peterson, Trenton M Tovar, Seth M Cohen
A high-throughput screening (HTS) method was devised to increase the rate of discovery and evaluation of nerve agent degradation catalysts. Using this HTS method, >90 solid state materials, predominantly metal-organic frameworks (MOFs), were analyzed for their ability to hydrolyze the nerve agent simulant methyl paraoxon at two pH values (8.0 and 10.0).
May 21, 2018: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/29780522/a-single-electrode-electrochemical-system-for-multiplex-electrochemiluminescence-analysis-based-on-a-resistance-induced-potential-difference
#16
Wenyue Gao, Kateryna Muzyka, Xiangui Ma, Baohua Lou, Guobao Xu
Developing low-cost and simple electrochemical systems is becoming increasingly important but still challenged for multiplex experiments. Here we report a single-electrode electrochemical system (SEES) using only one electrode not only for a single experiment but also for multiplex experiments based on a resistance induced potential difference. SEESs for a single experiment and multiplex experiments are fabricated by attaching a self-adhesive label with a hole and multiple holes onto an ITO electrode, respectively...
April 28, 2018: Chemical Science
https://www.readbyqxmd.com/read/29780503/a-high-throughput-screening-method-for-the-nano-crystallization-of-salts-of-organic-cations
#17
Philipp P Nievergelt, Martin Babor, Jan Čejka, Bernhard Spingler
The generation of solid salts of organic molecules is important to the chemical and pharmaceutical industry. Commonly used salt screening methods consume a lot of resources. We employed a combination of ion exchange screening and vapour diffusion for crystallization. This technique is semi-automatic and requires just nanoliters of the solution of the analyte to be crystallized. This high throughput screening yielded single crystals of sufficient size and quality for single-crystal X-ray structure determination using an in-house X-ray diffractometer...
April 21, 2018: Chemical Science
https://www.readbyqxmd.com/read/29779890/high-throughput-screening-enhances-kidney-organoid-differentiation-from-human-pluripotent-stem-cells-and-enables-automated-multidimensional-phenotyping
#18
Stefan M Czerniecki, Nelly M Cruz, Jennifer L Harder, Rajasree Menon, James Annis, Edgar A Otto, Ramila E Gulieva, Laura V Islas, Yong Kyun Kim, Linh M Tran, Timothy J Martins, Jeffrey W Pippin, Hongxia Fu, Matthias Kretzler, Stuart J Shankland, Jonathan Himmelfarb, Randall T Moon, Neal Paragas, Benjamin S Freedman
Organoids derived from human pluripotent stem cells are a potentially powerful tool for high-throughput screening (HTS), but the complexity of organoid cultures poses a significant challenge for miniaturization and automation. Here, we present a fully automated, HTS-compatible platform for enhanced differentiation and phenotyping of human kidney organoids. The entire 21-day protocol, from plating to differentiation to analysis, can be performed automatically by liquid-handling robots, or alternatively by manual pipetting...
May 15, 2018: Cell Stem Cell
https://www.readbyqxmd.com/read/29779767/novel-red-fluorescence-protein-based-microplate-assay-for-drug-screening-against-dormant-mycobacterium-tuberculosis-by-using-paraffin
#19
Amar Yeware, Dhiman Sarkar
The hypoxia model of dormancy is widely used in drug screening programs to identify novel inhibitors against latent Mycobacterium tuberculosis disease. In earlier reported microplate assays, hypoxia was maintained by either sealing the microplate or shifting in an anaerobic chamber to develop dormant phenotype. In these assays, inhibitors were added during inoculation, which mainly represents the active stage inhibitors instead of the dormant ones. Herein, the culture was covered with paraffin to develop hypoxia condition and consequently providing the advantage of adding compounds at any stage during incubation of 96-well plate...
May 2018: Tuberculosis
https://www.readbyqxmd.com/read/29779372/hollow-micropillar-array-method-for-high-capacity-drug-screening-on-filter-grown-epithelial-cells
#20
Byung-Ju Jin, Sujin Lee, Alan S Verkman
New high-throughput assay formats and innovative screening technologies are needed for miniaturized screens using small quantities of near-native, patient-derived cells. Here, we developed a hollow micropillar array method to screen compounds using epithelial cells cultured on a porous support, with the goal of screening thousands of compounds using a single 24-mm diameter transwell filter containing cultured cells. Test compounds (~1 nL) in an alginate hydrogel were printed by microinjection in hollow cylindrical micropillars (height 150 μm, inner diameter 100 μm) spaced 300 μm apart in a square array configuration...
May 19, 2018: Analytical Chemistry
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