Automated high-throughput dense matrix protein folding screen using a liquid handling robot combined with microfluidic capillary electrophoresis

2016 ◽  
Vol 120 ◽  
pp. 138-147 ◽  
Author(s):  
Philip An ◽  
Dwight Winters ◽  
Kenneth W. Walker
2005 ◽  
Vol 42 (6) ◽  
pp. 1063-1067 ◽  
Author(s):  
John R. Moriarity ◽  
Amanda D. Loftis ◽  
Gregory A. Dasch

2021 ◽  
Author(s):  
Katja Hellendahl ◽  
Maryke Fehlau ◽  
Sebastian Hans ◽  
Peter Neubauer ◽  
Anke Kurreck

Nucleoside kinases (NKs) are key enzymes involved in the in vivo phosphorylation of nucleoside analogues used as drugs to treat cancer or viral infections. Having different specificities, the characterization of NKs is essential for drug design and the production of nucleotide analogues in an in vitro enzymatic process. Therefore, a fast and reliable substrate screening assay for NKs is of great importance. Here, we report the validation of a well-known luciferase-based assay for the detection of NK activity in 96-well plate format. The assay was semi-automated using a liquid handling robot. A good linearity was demonstrated (r² >0.98) in the range of 0 to 500 µM ATP, and it was shown that also alternative phosphate donors like dATP or CTP were accepted by the luciferase. The developed high-throughput assay revealed comparable results to HPLC analysis. The assay was exemplary used for the comparison of the substrate spectra of four nucleoside kinases using 20 (8 natural and 12 modified) substrates. The screening results correlated well with literature data and, additionally, previously unknown substrates were identified for three of the NKs studied. Our results demonstrate that the developed semi-automated high-throughput assay is suitable to identify best performing NKs for a wide range of substrates.


2020 ◽  
Author(s):  
Tim Cernak ◽  
Babak Mahjour

<p>High throughput experimentation (HTE) is an increasingly important tool in the study of chemical synthesis. While the hardware for running HTE in the synthesis lab has evolved significantly in recent years, there remains a need for software solutions to navigate data rich experiments. We have developed the software, phactor™, to facilitate the performance and analysis of HTE in a chemical laboratory. phactor™ allows experimentalists to rapidly design arrays of chemical reactions in 24, 96, 384, or 1,536 wellplates. Users can access online reagent data, such as a lab inventory, to populate wells with experiments and produce instructions to perform the screen manually, or with the assistance of a liquid handling robot. After completion of the screen, analytical results can be uploaded for facile evaluation, and to guide the next series of experiments. All chemical data, metadata, and results are stored in a machine-readable format.</p>


2020 ◽  
Author(s):  
Tim Cernak ◽  
Babak Mahjour

<p>High throughput experimentation (HTE) is an increasingly important tool in the study of chemical synthesis. While the hardware for running HTE in the synthesis lab has evolved significantly in recent years, there remains a need for software solutions to navigate data rich experiments. We have developed the software, phactor™, to facilitate the performance and analysis of HTE in a chemical laboratory. phactor™ allows experimentalists to rapidly design arrays of chemical reactions in 24, 96, 384, or 1,536 wellplates. Users can access online reagent data, such as a lab inventory, to populate wells with experiments and produce instructions to perform the screen manually, or with the assistance of a liquid handling robot. After completion of the screen, analytical results can be uploaded for facile evaluation, and to guide the next series of experiments. All chemical data, metadata, and results are stored in a machine-readable format.</p>


2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Roman Jansen ◽  
Kira Küsters ◽  
Holger Morschett ◽  
Wolfgang Wiechert ◽  
Marco Oldiges

Abstract Background Morphology, being one of the key factors influencing productivity of filamentous fungi, is of great interest during bioprocess development. With increasing demand of high-throughput phenotyping technologies for fungi due to the emergence of novel time-efficient genetic engineering technologies, workflows for automated liquid handling combined with high-throughput morphology analysis have to be developed. Results In this study, a protocol allowing for 48 parallel microbioreactor cultivations of Aspergillus carbonarius with non-invasive online signals of backscatter and dissolved oxygen was established. To handle the increased cultivation throughput, the utilized microbioreactor is integrated into a liquid handling platform. During cultivation of filamentous fungi, cell suspensions result in either viscous broths or form pellets with varying size throughout the process. Therefore, tailor-made liquid handling parameters such as aspiration/dispense height, velocity and mixing steps were optimized and validated. Development and utilization of a novel injection station enabled a workflow, where biomass samples are automatically transferred into a flow through chamber fixed under a light microscope. In combination with an automated image analysis concept, this enabled an automated morphology analysis pipeline. The workflow was tested in a first application study, where the projected biomass area was determined at two different cultivation temperatures and compared to the microbioreactor online signals. Conclusions A novel and robust workflow starting from microbioreactor cultivation, automated sample harvest and processing via liquid handling robots up to automated morphology analysis was developed. This protocol enables the determination of projected biomass areas for filamentous fungi in an automated and high-throughput manner. This measurement of morphology can be applied to describe overall pellet size distribution and heterogeneity.


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