scholarly journals Simplified high-throughput methods for deep proteome analysis on the timsTOF Pro

2019 ◽  
Author(s):  
Jarrod J Sandow ◽  
Giuseppe Infusini ◽  
Laura F Dagley ◽  
Rune Larsen ◽  
Andrew I Webb
Keyword(s):  
High Throughput ◽  
Biological Samples ◽  
High Performance ◽  
Dynamic Range ◽  
Proteome Analysis ◽  
Single Shot ◽  
Custom Made ◽  
Mass Spectrometry Technology ◽  
Rapid Generation ◽  
Dependent Sample

AbstractRecent advances in mass spectrometry technology have seen remarkable increases in proteomic sequencing speed, while improvements to dynamic range have remained limited. An exemplar of this is the new timsTOF Pro instrument, which thanks to its trapped ion mobility, pushes effective fragmentation rates beyond 100Hz and provides accurate CCS values as well as impressive sensitivity. Established data dependent methodologies underutilize these advances by relying on long analytical columns and extended LC gradients to achieve comprehensive proteome coverage from biological samples. Here we describe the implementation of methods for short packed emitter columns that fully utilize instrument speed and CCS values by combining rapid generation of deep peptide libraries with enhanced matching of single shot data dependent sample analysis. Impressively, with only a 17 minute gradient separation (50 samples per day), the combination of high performance chromatography and CCS enhanced library based matching resulted in an average of 6,690 protein identifications within individual samples, and 7,797 proteins cumulatively across replicates from HeLa cell tryptic digests. Additionally, an ultra-high throughput setup utilizing 5 min gradients (180 samples per day) yielded an average of 2,800 protein identifications within individual samples and 4,254 proteins cumulatively across replicates. These workflows are simple to implement on available technology and do not require complex software solutions or custom made consumables to achieve high throughput and deep proteome analysis from biological samples.

scholarly journals Novel Electrochemiluminescent Assays for Drug Discovery

2000 ◽  
Vol 5 (1) ◽  
pp. 45-48 ◽  
Author(s):  
Maura C. Kibbey ◽  
David MacAllan ◽  
James W. Karaszkiewicz
Keyword(s):  
Drug Discovery ◽  
High Precision ◽  
High Throughput ◽  
High Performance ◽  
Dynamic Range ◽  
Wide Dynamic Range ◽  
Receptor Ligand ◽  
Enzymatic Assays ◽  
Detection Systems ◽  
Protein Assays

IGEN's ORIGEN® technology, which is based on electrochemiluminescence, has been adopted by a number of research and bioanalytical laboratories who have recognized its exquisite sensitivity, high precision, wide dynamic range, and flexibility in formatting a wide variety of applications. IGEN's M-SERIES™ marks the introduction of the second generation of detection systems employing the ORIGEN technology specifically repackaged to address the needs of the high throughput laboratories involved in drug discovery. Assays are formatted without wash steps. Users realize the high performance of a heterogeneous technology with the convenience of a homogeneous format. The M-SERIES platform can address enzymatic assays (kinases, proteases, helicases, etc.), receptor-ligand or protein-protein assays, immunoassays, quantitation of nucleic acids, as well as other applications. Recent assay formats will be explored in detail.

scholarly journals High-performance magneto-rheological clutches for direct-drive actuation: Design and development

2021 ◽  
pp. 1045389X2110069
Author(s):  
Sergey Pisetskiy ◽  
Mehrdad Kermani
Keyword(s):  
High Performance ◽  
Degrees Of Freedom ◽  
Dynamic Range ◽  
Complete Analysis ◽  
Mass Ratio ◽  
Direct Drive ◽  
Element Analysis ◽  
Prototype Development ◽  
Magneto Rheological ◽  
Hall Sensors

This paper presents an improved design, complete analysis, and prototype development of high torque-to-mass ratio Magneto-Rheological (MR) clutches. The proposed MR clutches are intended as the main actuation mechanism of a robotic manipulator with five degrees of freedom. Multiple steps to increase the toque-to-mass ratio of the clutch are evaluated and implemented in one design. First, we focus on the Hall sensors’ configuration. Our proposed MR clutches feature embedded Hall sensors for the indirect torque measurement. A new arrangement of the sensors with no effect on the magnetic reluctance of the clutch is presented. Second, we improve the magnetization of the MR clutch. We utilize a new hybrid design that features a combination of an electromagnetic coil and a permanent magnet for improved torque-to-mass ratio. Third, the gap size reduction in the hybrid MR clutch is introduced and the effect of such reduction on maximum torque and the dynamic range of MR clutch is investigated. Finally, the design for a pair of MR clutches with a shared magnetic core for antagonistic actuation of the robot joint is presented and experimentally validated. The details of each approach are discussed and the results of the finite element analysis are used to highlight the required engineering steps and to demonstrate the improvements achieved. Using the proposed design, several prototypes of the MR clutch with various torque capacities ranging from 15 to 200 N·m are developed, assembled, and tested. The experimental results demonstrate the performance of the proposed design and validate the accuracy of the analysis used for the development.

Fully automated sample treatment method for high throughput proteome analysis

2020 ◽  
Author(s):  
Huiming Yuan ◽  
Zhongpeng Dai ◽  
Xiaodan Zhang ◽  
Baofeng Zhao ◽  
Hongwei Chu ◽  
...  
Keyword(s):  
High Throughput ◽  
Proteome Analysis ◽  
Treatment Method ◽  
Sample Treatment

Directed Development of High-Performance Membranes via High-Throughput and Combinatorial Strategies

2006 ◽  
Vol 8 (2) ◽  
pp. 168-173 ◽  
Author(s):  
Metin Bulut ◽  
Lieven E. M. Gevers ◽  
Johan S. Paul ◽  
Ivo F. J. Vankelecom ◽  
Pierre A. Jacobs
Keyword(s):  
High Throughput ◽  
High Performance ◽  
Directed Development

scholarly journals Microplate Orbital Mixing Improves High-Throughput Cell-Based Reporter Assay Readouts

2006 ◽  
Vol 12 (1) ◽  
pp. 140-144 ◽  
Author(s):  
Michael K. Hancock ◽  
Myleen N. Medina ◽  
Brendan M. Smith ◽  
Anthony P. Orth
Keyword(s):  
High Throughput ◽  
Dynamic Range ◽  
Cell Lysis ◽  
Single Step ◽  
Luciferase Assay ◽  
Luciferase Reporter ◽  
Activity Measurements ◽  
Simple Detection ◽  
Reporter Assays ◽  
Orbital Mixing

Reporter assays are commonly used for high-throughput cell-based screening of compounds, cDNAs, and siRNAs due to robust signal, ease of miniaturization, and simple detection and analysis. Among the most widely used reporter genes is the bioluminescent enzyme luciferase, which, when exposed to its substrate luciferin upon cell lysis, yields linear signal over a dynamic range of several orders of magnitude. Commercially available luciferase assay formulations have been developed permitting homogeneous, single-step cell lysis and reporter activity measurements. Assay conditions employed with these formulations are typically designed to minimize well-to-well luminescence variability due to variability in dispensing, evaporation, and incomplete sample mixing. The authors demonstrate that incorporating a microplate orbital mixing step into 96- and 384-well microplate cell-based luciferase reporter assays can greatly improve reporter readouts. They have found that orbital mixing using commercially available mixers facilitates maximal luciferase signal generation from high cell density–containing samples while minimizing variability due to partial cell lysis, thereby improving assay precision. The authors fully expect that widespread availability of mixers with sufficiently small orbits and higher speed settings will permit gains in signal and precision in the 1536-well format as well.

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