scholarly journals SpeedScreen: The “Missing Link” between Genomics and Lead Discovery

2004 ◽  
Vol 9 (6) ◽  
pp. 498-505 ◽  
Author(s):  
Hartmut Zehender ◽  
Francois Le Goff ◽  
Natalie Lehmann ◽  
Ireos Filipuzzi ◽  
Lorenz M. Mayr
Keyword(s):  
High Throughput Screening ◽  
Assay Development ◽  
Size Exclusion ◽  
Ionization Mass ◽  
Label Free ◽  
Lead Discovery ◽  
Affinity Selection ◽  
Exclusion Chromatography ◽  
Microbore Liquid Chromatography ◽  
Selection Methodology

SpeedScreen is a novel, label-free, in-solution, affinity-based selection methodology for high-throughput screening (HTS) developed at Novartis Pharma. The SpeedScreen protocol comprises in-solution affinity selection, followed by size exclusion chromatography in combination with microbore-liquid-chromatography/electrospray-ionization mass spectrometry (micro-LC/ESI-MS). The authors describe the basic concept behind assay development, HTS, and data analysis with the SpeedScreen technology. Advantages and limitations of SpeedScreen compared to alternative screening technologies are discussed, and an example is given from a SpeedScreen campaign applying this innovative affinity selection concept in HTS.

scholarly journals A Chemoinformatics Analysis of Hit Lists Obtained from High-Throughput Affinity-Selection Screening

2005 ◽  
Vol 11 (2) ◽  
pp. 123-130 ◽  
Author(s):  
Nathan Brown ◽  
Hartmut Zehender ◽  
Kamal Azzaoui ◽  
Ansgar Schuffenhauer ◽  
Lorenz M. Mayr ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Size Exclusion ◽  
Label Free ◽  
Affinity Selection ◽  
High Performing ◽  
Exclusion Chromatography ◽  
Screening Platform ◽  
Screening Technology

The high-throughput affinity-selection screening platform SpeedScreen was recently reported by the Novartis Institutes for BioMedical Research as a homogeneous, label-free screening technology with mass-spectrometry readout. SpeedScreen relies on the screening of compound mixtures with various target proteins and uses fast size-exclusion chromatography to separate target-bound from unbound substances. After disintegration of the target-binder complex, the binder molecules are identified by their molecular masses using liquid chromatography/mass spectrometry. The authors report an analysis of the molecular properties of hits obtained with SpeedScreen on 26 targets screened within the past few years at Novartis using this technology. Affinity-based SpeedScreen is a robust high-throughput screening technology that does not accumulate frequent hitters or potential covalent binders. The hits are representative of the most commonly identified scaffold classes observed for known drugs. Validated SpeedScreen hits tend to be enriched on more lipophilic and larger-molecular-weight compounds compared to the whole library. The potential for a reduced SpeedScreen screening set to be used in case only limited protein quantities are available is evaluated. Such a reduced compound set should also maximize the coverage of the high-performing regions of the chemical property and class spaces; chemoinformatics methods including genetic algorithms and divisive K-means clustering are used for this aim.

scholarly journals Flurbiprofen: A Study of the Behavior of the Scalemate by Chromatography, Sublimation, and NMR

Symmetry ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 543
Author(s):  
Magdalena Kwiatkowska ◽  
Alicja Wzorek ◽  
Anna Kolbus ◽  
Mariusz Urbaniak ◽  
Jianlin Han ◽  
...  
Keyword(s):  
Relaxation Time ◽  
Preparative Thin Layer Chromatography ◽  
Transverse Relaxation Time ◽  
Size Exclusion ◽  
Test Case ◽  
Ionization Mass ◽  
Chiral Drugs ◽  
Good Test ◽  
Exclusion Chromatography ◽  
Medium Pressure Liquid Chromatography

2-(2-Fluoro-4-biphenyl) propionic acid (flurbiprofen), from the phenylalkanoic acid family of nonsteroidal anti-inflammatory drugs (NSAID’s), is currently on the pharmaceutical market as a racemate. This racemic compound was tested for its propensity to undergo the self-disproportionation of enantiomers (SDE) phenomenon by various forms of chromatography (SDEvC), such as routine gravity-driven column chromatography, medium-pressure liquid chromatography (MPLC), preparative thin-layer chromatography (PTLC), and size-exclusion chromatography (SEC), as well as by sublimation (SDEvS). Furthermore, examination by nuclear magnetic resonance (NMR) in various solvents found that flurbiprofen exhibited the phenomenon of self-induced diastereomeric anisochronism (SIDA). By measurement of the diffusion coefficient (D), the longitudinal relaxation time (T1), and the transverse relaxation time (T2) using NMR, as well as by electrospray ionization-mass spectrometry (ESI-MS) examinations, the preferred intermolecular association was found to be solvent dependent, e.g., heterochiral association was preferred in toluene, while homochiral association was preferred in more polar solvents. This study also attempted, unsuccessfully, to correlate the NMR measurements of flurbiprofen with chromatographic outcomes for the rationalization and prediction of chromatographic results based on NMR measurements. Because the intermolecular hydrogen bonding of the acid groups in flurbiprofen overwhelmingly predominates over other intermolecular interactions, flurbiprofen seemed to represent a good test case for this idea. The behavior of scalemic samples of flurbiprofen is important, as, although it is currently dispensed as a racemate, clinical applications of the R enantiomer have been investigated. SDEvC and SDEvS both have ramifications for the preparation, handling, and storage of enantioenriched flurbiprofen, and this concern applies to other chiral drugs as well.

scholarly journals Antibacterial Peptides Produced by Alcalase from Cowpea Seed Proteins

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 870
Author(s):  
Ali Osman ◽  
Gamal Enan ◽  
Abdul-Raouf Al-Mohammadi ◽  
Seham Abdel-Shafi ◽  
Samar Abdel-Hameid ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Seed Proteins ◽  
Negative Ions ◽  
Size Exclusion ◽  
Ionization Mass ◽  
Bacterial Cells ◽  
Antibacterial Efficiency ◽  
Exclusion Chromatography ◽  
Molecular Masses ◽  
Cowpea Seed

Cowpea seed protein hydrolysates (CPH) were output from cowpea seeds applying alcalase® from Bacillus licheniformis. CPH with an elevated level of hydrolysis was fractionated by size exclusion chromatography (SEC). Both CPH and SEC-portions showed to contain antimicrobial peptides (AMPs) as they inhibited both Gram-positive bacteria, such as Listeria monocytogenes LMG10470 (L. monocytogenes), Listeria innocua. LMG11387 (L. innocua), Staphylococcus aureus ATCC25923 (S.aureus), and Streptococcus pyogenes ATCC19615 (St.pyogenes), and Gram-negative bacteria, such as Klebsiella pnemoniae ATCC43816 (K. pnemoniae), Pseudomonas aeroginosa ATCC26853 (P. aeroginosa), Escherichia coli ATCC25468) (E.coli) and Salmonella typhimurium ATCC14028 (S. typhimurium).The data exhibited that both CPH and size exclusion chromatography-fraction 1 (SEC-F1) showed high antibacterial efficiency versus almost all the assessed bacteria. The MIC of the AMPs within SEC-F1 and CPHs were (25 µg/mL) against P. aeruginosa, E.coli and St. pyogenes. However, higher MICsof approximately 100–150 µg/mL showed for both CPHs and SEC-F1 against both S. aureus and L. innocua; it was 50 µg/mL of CPH against S.aureus. The Electro-spray-ionization-mass-spectrometry (ESI-MS) of fraction (1) revealed 10 dipeptides with a molecular masses arranged from 184 Da to 364 Da and one Penta peptide with a molecular mass of approximately 659 Da inthe case of positive ions. While the negative ions showed 4 dipeptides with the molecular masses that arranged from 330 Da to 373 Da. Transmission electron microscope (TEM) demonstrated that the SEC-F1 induced changes in the bacterial cells affected. Thus, the results suggested that the hydrolysis of cowpea seed proteins by Alcalase is an uncomplicated appliance to intensify its antibacterial efficiency.

High-throughput screening of the chemical composition distribution of random styrene-butyl acrylate copolymers

e-Polymers ◽  
2005 ◽  
Vol 5 (1) ◽  
Author(s):  
Iván García Romero ◽  
Harald Pasch
Keyword(s):  
Chemical Composition ◽  
High Throughput ◽  
Butyl Acrylate ◽  
High Throughput Screening ◽  
Size Exclusion ◽  
Chemical Compositions ◽  
Composition Distribution ◽  
Chemical Composition Distribution ◽  
Exclusion Chromatography ◽  
Liquid Chromatographic

AbstractThe development of high-throughput liquid chromatographic techniques for the analysis of styrene-butyl acrylate (SBA) copolymers is discussed. The analysis time in size-exclusion chromatography (SEC) can be reduced to about 3 min per sample when high-throughput SEC columns and high flow rates are used. In gradient HPLC, small columns with improved separation efficiencies can be applied. The time requirements can be decreased to less than 2 min per sample. Using the high-throughput HPLC technique, the chemical composition distribution of high-conversion SBA copolymers can be analyzed in a fast and efficient way. The calibration of HPLC separation is conducted by coupling the HPLC system with FTIR through the LC-transform interface. A comparison of the chemical compositions of the copolymers obtained by 1H NMR, off-line FTIR and coupled HPLCFTIR verifies the accuracy of the high-throughput copolymer analysis approach.

scholarly journals MALDIViz: A Comprehensive Informatics Tool for MALDI-MS Data Visualization and Analysis

2017 ◽  
Vol 22 (10) ◽  
pp. 1246-1252 ◽  
Author(s):  
Kishore Kumar Jagadeesan ◽  
Simon Ekström
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Web Application ◽  
Low Cost ◽  
Maldi Ms ◽  
Ionization Mass ◽  
Label Free ◽  
Label Free Detection ◽  
R Shiny

Recently, mass spectrometry (MS) has emerged as an important tool for high-throughput screening (HTS) providing a direct and label-free detection method, complementing traditional fluorescent and colorimetric methodologies. Among the various MS techniques used for HTS, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) provides many of the characteristics required for high-throughput analyses, such as low cost, speed, and automation. However, visualization and analysis of the large datasets generated by HTS MALDI-MS can pose significant challenges, especially for multiparametric experiments. The datasets can be generated fast, and the complexity of the experimental data (e.g., screening many different sorbent phases, the sorbent mass, and the load, wash, and elution conditions) makes manual data analysis difficult. To address these challenges, a comprehensive informatics tool called MALDIViz was developed. This tool is an R-Shiny-based web application, accessible independently of the operating system and without the need to install any program locally. It has been designed to facilitate easy analysis and visualization of MALDI-MS datasets, comparison of multiplex experiments, and export of the analysis results to high-quality images.

scholarly journals Optimization of Unnickedβ2-Glycoprotein I and High Avidity Anti-β2-Glycoprotein I Antibodies Isolation

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Andrej Artenjak ◽  
Adrijana Leonardi ◽  
Igor Križaj ◽  
Aleš Ambrožič ◽  
Snezna Sodin-Semrl ◽  
...  
Keyword(s):  
Isolation Procedure ◽  
Size Exclusion ◽  
Ionization Mass ◽  
High Avidity ◽  
Isolation And Purification ◽  
Average Mass ◽  
Glycoprotein I ◽  
Unknown Protein ◽  
Exclusion Chromatography ◽  
Weight Protein

Patient biological material for isolation ofβ2-glycoprotein I (β2GPI) and high avidity IgG anti-β2-glycoprotein I antibodies (HAv anti-β2GPI) dictates its full utilization. The aim of our study was to evaluate/improve procedures for isolation of unnickedβ2GPI and HAv aβ2GPI to gain unmodified proteins in higher yields/purity. Isolation ofβ2GPI from plasma was a stepwise procedure combining nonspecific and specific methods. For isolation of polyclonal HAv aβ2GPI affinity chromatographies with immobilized protein G and humanβ2GPI were used. The unknown protein found during isolation was identified by liquid chromatography electrospray ionization mass spectrometry and the nonredundant National Center for Biotechnology Information database. The average mass of the isolated unnicked purifiedβ2GPI increased from 6.56 mg to 9.94 mg. In the optimized isolation procedure the high molecular weight protein (proteoglycan 4) was successfully separated fromβ2GPI in the 1st peaks with size exclusion chromatography. The average efficiency of the isolation procedure for polyclonal HAv anti-β2GPI from different matrixes was 13.8%, as determined by ourin-houseanti-β2GPI ELISA. We modified thein-houseisolation and purification procedures of unnickedβ2GPI and HAv anti-β2GPI, improving the purity of antigen and antibodies as well as increasing the number of tests routinely performed with thein-houseELISA by ~50%.

Molecular Weight and Tacticity of Oligoacrylates by Capillary Electrophoresis - Mass Spectrometry

2010 ◽  
Vol 63 (8) ◽  
pp. 1219 ◽  
Author(s):  
Marianne Gaborieau ◽  
Tim J. Causon ◽  
Yohann Guillaneuf ◽  
Emily F. Hilder ◽  
Patrice Castignolles
Keyword(s):  
Capillary Electrophoresis ◽  
Raft Polymerization ◽  
Degree Of Polymerization ◽  
Size Exclusion ◽  
Separation Mechanism ◽  
Ionization Mass ◽  
Polymeric Particles ◽  
Reversible Addition ◽  
Exclusion Chromatography

Oligo(acrylic acid) efficiently stabilizes polymeric particles, especially particles produced by reversible addition–fragmentation chain transfer (RAFT) (as hydrophilic block of an amphiphilic copolymer). Capillary electrophoresis (CE) has a far higher resolution power to separate these oligomers than the commonly used size exclusion chromatography. Coupling CE to electrospray ionization mass spectrometric detection unravels the separation mechanism. CE separates these oligomers, not only according to their degree of polymerization, but also according to their tacticity, in agreement with NMR analysis. Such analysis will provide insight into the role of these oligomers as stabilizers in emulsion polymerization, and into the mechanism of the RAFT polymerization with respect to degree of polymerization and tacticity.

Sign in / Sign up

Export Citation Format

Share Document